Stable liquid formulations of glucagon-like peptide 1 or analogues thereof

ABSTRACT

Stable liquid formulations of GLP-1 and GLP-1 analogues and method of using such formulations in the treatment of disorders or conditions are provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to GB 1917723.7, filed Dec. 4, 2019.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jan. 4, 2021, isnamed 126672-0102_SL.txt and is 2,878 bytes in size.

TECHNICAL FIELD

The present disclosure relates to formulations of GLP-1 and GLP-1analogues thereof and method of using such formulations.

BACKGROUND

The native glucagon-like peptide 1 (native GLP-1) is a 37 amino acidpeptide that is secreted by the L-cells of the intestine in response tofood ingestion. It has been found to stimulate insulin secretion,thereby causing glucose uptake by cells and a decrease in serum glucoselevels (Mojsov, S., 1992). Native GLP-1 is susceptible to proteolyticcleavage which gives rise to two biologically active peptides;GLP-1(7-37) and GLP-1(7-36)NH₂.

However, these biologically active peptides have a very short half-life(<2 minutes), due to their rapid metabolic degradation by the ubiquitousdipeptidyl dipeptidase-4 (DPP-4).

ROSE-010 (Val8-GLP-1) is a GLP-1 analogue that has been made resistantto DPP-4-mediated cleavage by replacement of the alanine residue atposition 8 with valine. ROSE-010 has a very high level of sequenceidentity with the native GLP-1(7-37) peptide and is a potent receptoragonist.

In clinical trials, ROSE-010 has been shown to reduce acuteexacerbations of irritable bowel syndrome (IBS) (Hellström P M, et al.,2009). In constipation predominant IBS (IBS-C), ROSE-010 delayed gastricemptying of solids; the accelerated colonic transit at 48 h with 30 and100 μg of ROSE-010 suggests potential for relief of constipation inIBS-C(Camilleri, M et al., 2012).

However, ROSE-010 forms noncovalent aggregates in solution (Doyle, B. D.et al, 2005) that renders the development of liquid formulationschallenging.

Stable liquid formulations of GLP-1 and GLP-1 analogues such as ROSE-010are described herein.

SUMMARY

Described herein are liquid formulations of glucagon-like peptide-1 orof GLP-1 analogues.

In certain aspects and embodiments described herein, the liquidpharmaceutical formulations of the present disclosure may include GLP-1or GLP-1 analogues that are agonists of the GLP-1 receptor.Pharmaceutical formulations of glucagon-like peptide-1 (GLP-1 (7-37)) orof GLP-1 analogue such as ROSE-010 thereof are provided.

In other aspects and embodiments as described herein, the pharmaceuticalformulations may be aqueous and may include ROSE-010 in solution. Thepharmaceutical formulations of the present disclosure may advantageouslywithstand long-term storage. For example, the pharmaceuticalformulations of the present disclosure may advantageously have a minimumstable shelf life of at least three months or more.

The pharmaceutical formulations of the present disclosure mayadvantageously withstand physical stress such as freeze-thaw, mechanicalstress and/or elevated temperature. The pharmaceutical formulation ofthe present disclosure may be suitable for injection. In certain aspectsand embodiments described herein the pharmaceutical formulations may beprovided in pre-filled syringes. In other aspects and embodimentsdescribed herein, the pharmaceutical formulations may be provided insingle-dose or multiple-dose containers.

In aspects and embodiments described herein, the pharmaceuticalformulations may be aqueous and may comprise a GLP-1 or GLP-1 analoguein solution such as to obtain a liquid pharmaceutical formulation of theGLP-1 or GLP-1 analogue.

The GLP-1 or GLP-1 analogues of the present disclosure have, for exampleand without limitation, the amino acid sequence set forth in SEQ IDNO:1.

An exemplary embodiment of GLP-1 or GLP-1 analogues includes thebiologically active peptide GLP-1(7-37) which comprises the amino acidsequence set forth in SEQ ID NO: 2(H-HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG-OH). The GLP-1 analogues of thepresent disclosure may have one or more amino acid substitutions incomparison with the wild type GLP-1 peptide. The amino acidsubstitutions may include conservative amino acid substitutions. Theamino acid substitutions may include for example, modification of theamino acid residue 1, modification of amino acid residue 2, modificationof amino acid residue 15, modification of amino acid residue 21,modification of amino acid residue 31 and combination thereof.

Exemplary embodiments of GLP-1 analogues of the present disclosure mayinclude, for example and without limitation, a GLP-1 analogue where theC-terminal glycine residue is absent(H-HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR-NH₂ (GLP-1(7-36)NH₂, SEQ ID NO:4)).

Additional exemplary embodiments of analogues described herein mayinclude, for example, those where the first amino acid residue is amodified histidine residue such as for example and without limitation,L-histidine, D-histidine, desaminohistidine, 2-amino-histidine,beta-hydroxy-histidine, homohistidine, alpha-fluoromethylhistidine oralpha-methyl-histidine.

Other exemplary embodiments of analogues described herein may includethose where the second amino acid is alanine, valine, glycine,threonine, isoleucine, or alpha-methyl-Ala. The ROSE-010 peptide whichincludes a valine residue instead of alanine at the second position(H-HVEGTFTSDVSSYLEGQAAKEFIAWLVKGRG-OH (SEQ ID NO:3)) is encompassed bythe present disclosure. The C-terminal glycine residue may be absentfrom SEQ ID NO:3 so as to form a shorter peptide(H-HVEGTFTSDVSSYLEGQAAKEFIAWLVKGR-NH₂ (SEQ ID NO:5)).

Additional GLP-1 analogues of the present disclosure may include, forexample, those where the amino acid residue at position 15 (withreference to SEQ ID NO:2) is glutamic acid, glutamine, alanine,threonine, serine or glycine.

Additional GLP-1 analogues of the present disclosure may include, forexample, those where the amino acid residue at position 21 (withreference to SEQ ID NO:2) is glutamic acid, glutamine, alanine,threonine, serine or glycine.

Further GLP-1 analogues of the present disclosure may include, forexample, those where the amino acid residue at position 31 (withreference to SEQ ID NO:2) may be absent or is Gly-NH₂ or Gly-OH.

GLP-1 analogues of the present disclosure may also include, for example,those that have one or more of the amino acid substitutions exemplifiedherein or in the various substituents of SEQ ID NO:3.

The GLP-1 and GLP-1 analogues of the present disclosure may beformulated as stable liquid formulations or stable liquid pharmaceuticalformulations. The liquid pharmaceutical formulations may be water-based(i.e., aqueous formulations).

In an embodiment, the pharmaceutical formulations of the presentdisclosure may include, for example, the GLP-1(7-37) peptide (SEQ IDNO:2) or the GLP-1(7-36)NH₂ peptide (SEQ ID NO:4).

In another embodiment, the liquid pharmaceutical formulations of thepresent disclosure may include, for example, a GLP-1 analogue thatincludes or consists in the amino acid sequence set forth in SEQ IDNO:3.

In another exemplary embodiment, the GLP-1 analogues comprise an aminoacid sequence at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% identical to SEQ ID NO:3 wherein the amino acid at position 2is valine.

Other GLP-1 analogues have been described in U.S. Pat. Nos. 6,583,111 in8,642,548, WO2007/028394 and in WO91/11457, the entire content of whichis enclosed herein by reference.

The liquid pharmaceutical formulations of the present disclosure mayinclude, for example, and without limitation, a GLP-1 or a GLP-1analogue at a concentration ranging from about 10 μg/ml to about 1mg/ml, at a concentration ranging from about 50 μg/ml to about 750μg/ml, at a concentration ranging from about 100 μg/ml to about 500μg/ml, at a concentration ranging from about 200 μg/ml to about 500μg/ml, at a concentration of at least 300 μg/ml±20%, at a concentrationof 300 μg/ml±20%, at a concentration of 300 μg/ml±10% etc.

Exemplary embodiments of pharmaceutical formulations provided are thoseincluding ROSE-010 peptide (SEQ ID NO:3) at a concentration of, forexample, from about 10 μg/ml to about 1 mg/ml, from about 50 μg/ml toabout 750 μg/ml, from about 100 μg/ml to about 500 μg/ml, from about 200μg/ml to about 500 μg/ml, at a concentration of at least 300 μg/ml±10%,at a concentration of 300 μg/ml±10%, at a concentration of at least 300μg/ml±20%, at a concentration of 300 μg/ml±20% etc.

It is to be understood herein that in a given formulation, the peptideconcentration may vary over time depending on stress conditions to whichit is submitted. For an initial peptide concentration of 0.300 mg/ml, avariation of ±0.05 mg/ml (i.e., approximately ±17%) may be consideredwithin target range for quality control purposes. Higher variations maybe acceptable as certain conditions require. A peptide concentration of0.300±0.05 mg/ml, represents, for example, a range of 0.250 mg/ml to0.350 mg/ml.

It is to be understood herein that the expression “from about 10 μg/mlto about 1 mg/ml” includes any individual values (including fractions)comprised within and including 10 μg/ml and 1 mg/ml, such as forexample, 10 μg/ml, 15 μg/ml, 20 μg/ml, 30 μg/ml, 40 μg/ml, 50 μg/ml, 60μg/ml, 70 μg/ml, 80 μg/ml, 90 μg/ml, 100 μg/ml, 125 μg/ml, 150 μg/ml,175 μg/ml, 200 μg/ml, 225 μg/ml, 250 μg/ml, 275 μg/ml, 300 μg/ml, 325μg/ml, 350 μg/ml, 375 μg/ml, 400 μg/ml, 425 μg/ml, 450 μg/ml, 475 μg/ml,500 μg/ml, 525 μg/ml, 550 μg/ml, 575 μg/ml, 600 μg/ml, 625 μg/ml, 650μg/ml, 675 μg/ml, 700 μg/ml, 725 μg/ml, 750 μg/ml, 775 μg/ml, 800 μg/ml,825 μg/ml, 850 μg/ml, 875 μg/ml, 900 μg/ml, 925 μg/ml, 950 μg/ml, 975μg/ml and 1 mg/ml etc.

It is to be understood herein that the expression “from about 10 μg/mlto about 1 mg/ml” also includes any individual sub-ranges (includingfractions) comprised within and including from about 10 μg/ml to about 1mg/ml, such as for example, “from about 110 μg/ml to about 490 μg/ml”,“from about 205 μg/ml to about 999 μg/ml”, “from about 225 μg/ml toabout 875 μg/ml”, “from about 200 μg/ml to about 750 μg/ml” and thelike.

The same applies for similar expressions such as and not limited to“from about 200 μg/ml to about 500 μg/ml”, “from about 100 μg/ml toabout 500 μg/ml” and “from about 50 μg/ml to about 750 μg/ml” and thelike which comprise any individual values (and fractions) comprisedwithin and including such value and any individual sub-ranges (andfractions) comprised within and including such range.

In accordance with an aspect of the disclosure, stock solutions andformulations contain water. The type of water used in the preparation ofstock solutions and formulations may meet the guidance of the EuropeanPharmacopoeia (8th edition 2014) and may include for example, highlypurified water, ultrapure water and the like. In accordance with anembodiment of the disclosure the formulations may be prepared usingwater for injection.

In a first aspect provided is a liquid pharmaceutical formulation thatincludes a GLP-1 or GLP-1 analogue and a buffering agent.

The buffering agent described herein may be a single buffer. Thebuffering agent described herein may be suitable for injection in amammal (e.g., subcutaneous, intravenous, intradermal, intramuscular).The buffering agent of the present disclosure may be aqueous-based andmay be selected, for example, for its lack of adverse interaction withthe active ingredient. The buffering agent of the present disclosure mayallow the formulation to remain at a stable pH, especially at a pH rangeof 6.0 to 8.0 such as for example at a pH range of 6.0 to 7.5±0.15 andmore specifically within a range of 6.5 to 7.0±0.15. In an exemplaryembodiment the pH range is between 6.5±0.5 to 7.0±0.5.

It is to be understood herein that for a given formulation, the pH mayvary over time while remaining within an acceptable range. For example,the pH of a batch formulation may be set at an initial pH of 7.0 and thepH measured after 1 month, 3 months, 6 months storage may vary from 6.0to 8.0.

Moreover, although the target pH of a batch formulation may be set at7.0±0.5, a pH range of between 6.5±0.5 to 7.5±0.5 is understood hereinas being within specification.

Unless indicated otherwise, pH measurements are generally performed atroom temperature (e.g., generally between 20 and 25° C. and moreparticularly between 22-24° C.). The buffering agent described hereinmay include, for example and without limitation, acetate, carbonate,citrate, histidine, maleate, phosphate, succinate, tartrate,tromethamine and the like and combination thereof. Other bufferingagents may be suitable.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, acetate (e.g., sodium acetate) asbuffering agent.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, citrate (e.g., sodium citrate) asbuffering agent.

In additional embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, histidine (e.g., histidine-HCl)as buffering agent.

In additional embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, maleate (e.g., sodium maleate) asbuffering agent.

In further embodiments described herein, the pharmaceutical formulationsmay include, for example, phosphate as buffering agent. Exemplaryembodiments of phosphate buffer include without limitation sodiumphosphate, potassium phosphate, phosphate-buffered saline (PBS).

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, succinate as buffering agent.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, tartrate (e.g., potassium sodiumtartrate) as buffering agent.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, tromethamine as buffering agent.

In some embodiments of the present disclosure, the buffering agent maybe at a concentration ranging from about 1 mM to about 50 mM; from about2 mM to about 20 mM; from about 5 mM to about 20 mM; from about 5 mM toabout 15 mM; from about 7.5 mM to about 12.5 mM. The concentration ofthe buffering agent may be, for example, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM,10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM. However, it is to beunderstood that the concentration of the buffering agent can be eitherhigher or lower as specific conditions require.

It is to be understood herein that the expression “from about 1 mM toabout 50 mM” includes any individual values (including fractions)comprised within and including 1 mM and 50 mM, for example “from about 1mM to about 50 mM” includes 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mMand 50 mM.

It is to be understood herein that the expression “from about 1 mM toabout 50 mM” also includes any individual sub-ranges (includingfractions) comprised within and including “from about 1 mM to about 50mM”, such as for example, “from about 2 mM to about 50 mM”, “from about2 mM to about 49 mM”, “from about 5 mM to about 50 mM”, “from about 15mM to about 40 mM”, “from about 15 mM to about 20 mM”, “from about 15 mMto about 30 mM”, “from about 26 mM to about 30 mM”, “from about 32 mM toabout 49 mM” and the like.

The same applies for all similar expressions such as and not limited to“from about 0.5 mM to about 250 mM”, “from about 5 mM to about 15 mM”,“from about 7.5 mM to about 12.5 mM”, “from about 25 mM to about 75 mM”,“from about 50 mM to about 150 mM”, from about 165 mM to about 275 mM”,“from about 150 mM to about 300 mM” etc. which comprise any individualvalues (including fractions) comprised within and including such valueand any individual sub-ranges (including fractions) comprised within andincluding such range.

Exemplary embodiments of the pharmaceutical formulations describedherein, may include, for example and without limitation, bufferingagents at a concentration of about 10 mM±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include an acetate buffer (e.g., sodium acetate) at a concentrationof 10 mM±10%.

In other embodiments described herein, the pharmaceutical formulationsmay include a citrate buffer (e.g., sodium citrate) at a concentrationof 10 mM±10%.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include histidine buffer (e.g., histidine-HCl) at aconcentration of 10 mM±10%.

In further embodiments described herein, the pharmaceutical formulationsmay include, a maleate buffer (e.g., sodium maleate) at a concentrationof 10 mM±10%.

In other embodiments described herein, the pharmaceutical formulationsmay include a phosphate buffer (e.g., sodium phosphate) at aconcentration of 10 mM±10%.

In further embodiments described herein, the pharmaceutical formulationsmay include, a succinate buffer at a concentration of 10 mM±10%.

In additional embodiments described herein, the pharmaceuticalformulations may include, a tartrate buffer (e.g., potassium sodiumtartrate) at a concentration of 10 mM±10%.

In other embodiments described herein, the pharmaceutical formulationsmay include a tromethamine buffer at a concentration of 10 mM±10%.

In other aspects provided is a liquid pharmaceutical formulation thatincludes a GLP-1 or GLP-1 analogue, a buffering agent and a tonicityagent.

The tonicity agent may contribute to the osmolality of the solution. Theosmolality of the pharmaceutical formulations described herein may beadjusted to maximize the active ingredient's stability and/or tominimize discomfort to the patient upon administration. Tonicity agentsdescribed herein may render the formulation isotonic with blood and/orserum.

The osmolality of the pharmaceutical formulations of the presentdisclosure may vary, for example and without limitation, from 250-400mOsmol/kg; from about 275-375 mOsmol/kg; from about between 276-369mOsmol/kg; from about 290-370 mOsmol/kg. The osmolality of the liquidpharmaceutical formulations described herein may range from aboutbetween 275-299 mOsmol/kg (blood osmolality). However, it is to beunderstood that the osmolality can be either higher or lower as specificconditions require.

In some embodiments, the tonicity agent can include sugar-based tonicityagents such as sugars and polyols. Exemplary embodiments of basedtonicity agents include for example and without limitation, mannitol,sucrose, glucose, dextrose, trehalose and the like.

Exemplary embodiments of tonicity agents of the present disclosure mayinclude, for example and without limitation, dextrose, glucose, glyceringlycerol, mannitol, potassium chloride, sodium chloride, sodium sulfate,sorbitol, sucrose, trehalose and the like and combination thereof. Othertonicity agents may be suitable.

In some embodiments, the formulation may include glycerol. Theconcentration of glycerol may vary, for example and without limitation,from about 0.5% (v/v) to about 5% (v/v); from about 1% (v/v) to about 5%(v/v); from about 1.5% (v/v) to about 4.5% (v/v); from about 1% (v/v) toabout 5% (v/v); from about 2% (v/v) to about 4% (v/v); from about 1.5%(v/v) to about 4.5% (v/v); from about 1% (v/v) to about 5% (v/v) etc. Insome embodiments, the concentration of glycerol may vary, for exampleand without limitation, from about 2.5% (v/v) to about 3.5% (v/v). Inother embodiments, the concentration of glycerol may be, for example,2.5%±10% (or 275 mM±10%). However, it is to be understood that theconcentration of glycerol can be either higher or lower as specificconditions require.

In further embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, glycerolat a concentration of about 165 mM±10%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, glycerolat a concentration of about 275 mM±10%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include mannitol. The concentration of mannitol in theformulation may range, for example and without limitation, from about 10mM to about 500 mM, from about 50 mM to about 400 mM, from about 100 mMto about 400 mM; from about 150 mM to about 350 mM, from about 150 mM toabout 300 mM etc. In some embodiments, the concentration of mannitol mayrange from about 165 mM±10% to about 275 mM±10%. However, it is to beunderstood that the concentration of mannitol can be either higher orlower as specific conditions require. The concentration of mannitol mayalso be expressed herein in weight by volume (w/v) percentages.

In further embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, mannitolat a concentration of about 165 mM±10%. A concentration of 165 mM ofmannitol also corresponds to approximately 3%.

In additional embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, mannitolat a concentration of about 275 mM±10%. A concentration of 275 mM ofmannitol also corresponds to approximately 5%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include sodium chloride (NaCl). The concentration ofsodium chloride used in the formulation may be determined, for example,based on the desired method of administration. In some embodiments, theconcentration of sodium chloride may be for example at 0.9% (w/v)±10% soas to be isotonic to blood. In other embodiments, the concentration ofsodium chloride may vary, for example and without limitation, from about1 mM to about 250 mM; from about 10 mM to about 200 mM; from about 25 mMto about 200 mM; from about 40 mM to about 175 mM; from about 50 mM±10%to about 150 mM±10% etc. The concentration of sodium chloride may be forexample and without limitation, 50 mM±10%, 130 mM±10%, 150 mM±10% etc.However, it is to be understood that the concentration of sodiumchloride can be either higher or lower as specific conditions require.The concentration of sodium chloride may also be expressed herein inweight by volume (w/v) percentages.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sodiumchloride at a concentration of about 50 mM±10%. A concentration of 50 mMof NaCl also corresponds to approximately 0.3%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sodiumchloride at a concentration of about 130 mM±10%. A concentration of 130mM of NaCl also corresponds to approximately 0.7%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sodiumchloride at a concentration of about 150 mM±10%. A concentration of 150mM of NaCl also corresponds to approximately 0.9%.

In further embodiments of the present disclosure, the pharmaceuticalformulations may include, for example, sucrose. The concentration ofsucrose in the formulation is expressed in % of weight/volume (w/v) andmay range from about 1% (w/v) to 20% (w/v); from about 2% to 18%; fromabout 3% to 15%; from about 5% to 15%, etc. The concentration of sucrosein the formulation may be, for example and without limitation, 5%, 6%,7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% etc. In some embodiments, theconcentration of sucrose may range, for example, from about 8%±10% toabout 10%±10%. However, it is to be understood that the concentration ofsucrose can be either higher or lower as specific conditions require.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sucrose ata concentration of about 5%±10%.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sucrose ata concentration of about 8%±10%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, sucrose ata concentration of about 10%±10%.

In other embodiments, the formulation may include trehalose. Theconcentration of trehalose may vary, for example and without limitation,from about 5% (w/v) to about 25% (w/v); from about 7.5% (w/v) to about15% (w/v); from about 7.5% (w/v) to about 10% (w/v). In someembodiments, the concentration of trehalose may be about 10% (w/v)±10%.However, it is to be understood that the concentration of trehalose canbe either higher or lower as specific conditions require.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, trehaloseat a concentration of about 5%±10%.

In some embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, trehaloseat a concentration of about 8%±10%.

In other embodiments of the present disclosure, the pharmaceuticalformulations may include, for example and without limitation, trehaloseat a concentration of about 10%±10%.

It is to be understood herein that the expression “from about 5% toabout 15%” includes any individual values (including fractions)comprised within and including 5.0% and 15.0% such as for example 5%,5.25%, 5.5%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8%,8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 9.8%, 9.9%, 10%, 10.25%,10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%, 12.5%,12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 4.25%, 14.5%, 14.75%,14.8%, 15.0%.

It is to be understood herein that the expression “from about 5% toabout 15%” also includes any individual sub-ranges (including fractions)comprised within and including “from about 5.0% to about 15.0%”, “fromabout 5.0% to about 10.0%”, “from about 5.0% to about 8.0%”, “from about8.0% to about 10.0%”, “from about 6.0% to about 7.5%” and the like.

The same applies for all similar expressions such as and not limited to“from about 8% to about 10%” or “from about 0.01 to about 0.03%” etc.which comprise any individual values (and fractions) comprised withinand including such value and any individual sub-ranges (and fractions)comprised within and including such range. The expression “±10%” withrespect to a given value, is to be understood herein to encompass allindividual values such as +0%, +1%, +2%, +3%, +4%, +5%, +6%, +7%, +8%,+9%, +10%, −1%, −2%, −3%, −4%, −5%, −6%, −7%, −8%, −9%, −10%. Forexample, the expression “0.3 mg/ml±10%” includes any individual values(including fractions and ranges) comprised within 0.27 mg/ml and 0.33mg/ml.

The same applies for all similar expressions such as and not limited to“10% (v/v)±10%” etc. which comprises values such as 9%, 10% and 11% andany fractions or ranges in between such as 9.2%, 9.5%, 10.5% and thelike.

In other aspect provided, the pharmaceutical formulation may furtherinclude an excipient.

Excipients described herein include, for example, stabilizers,surfactants and combination thereof. However, it is to be understoodthat other excipients may be used.

Exemplary embodiments of stabilizers include, for example and withoutlimitations, amino acids (e.g., L-amino acids or D-amino acids). Aminoacid stabilizers include, for example and without limitations, alanine,arginine, glutamate, glycine, isoleucine, leucine, methionine, proline,and the like and combination thereof. Exemplary embodiments of aminoacid stabilizers include for example, L-alanine, L-arginine,L-glutamate, L-glycine, L-isoleucine, L-leucine, L-methionine,L-proline, etc.

The concentration of amino acids in the pharmaceutical formulationsdisclosed herein may vary from about 0.5 mM to about 300 mM; from about1 mM to about 200 mM; from about 5 mM to about 150 mM; from about 5 mMto about 100 mM; from about 7.5 mM to about 100 mM; from about 7.5 mM toabout 75 mM; from about 7.5 mM to about 50 mM; from about 10 mM to about50 mM etc. The concentration of amino acids in the pharmaceuticalformulations disclosed herein may be for example and withoutlimitations, 0.5 mM±10%, 1 mM±10%, 2 mM 10%, 3 mM 10%, 4 mM 10%, 5 mM10%, 10 mM±10%, 15 mM 10%, 20 mM±10%, 25 mM 10%, 30 mM±10%, 35 mM 10%,40 mM±10%, 45 mM 10% or 50 mM±10%. However, it is to be understood thatthe concentration of amino acids can be either higher or lower asspecific conditions require.

In certain embodiments of the present disclosure, the liquidpharmaceutical formulations may include alanine (e.g., L-alanine) at aconcentration of about 1 mM to about 250 mM±10%. In some embodiments ofthe present disclosure, the pharmaceutical formulations may includealanine at a concentration of about 5 mM to about 15 mM±10%. In otherembodiments of the present disclosure, the pharmaceutical formulationsmay include alanine at a concentration of about 10 mM±10%.

In other embodiments of the present disclosure, the liquidpharmaceutical formulations may include arginine (e.g., L-arginine) at aconcentration of from about 1 mM to about 250 mM±10%. In someembodiments of the present disclosure, the liquid pharmaceuticalformulations may include arginine at a concentration of about 100 mM±10%or at a concentration of 150 mM±10%.

In other embodiments of the present disclosure, the liquidpharmaceutical formulations may include glycine (e.g., L-glycine) at aconcentration of from about 1 mM to about 250 mM±10%. In someembodiments of the present disclosure, the liquid pharmaceuticalformulations may include glycine at a concentration of about 50 mM±10%or at a concentration of 150 mM±10%.

In other embodiments of the present disclosure, the liquidpharmaceutical formulations may include isoleucine (e.g., L-isoleucine)at a concentration of from about 1 mM to about 250 mM±10%. In someembodiments of the present disclosure, the liquid pharmaceuticalformulations may include isoleucine at a concentration of about 100mM±10% or at a concentration of 150 mM±10%.

In other embodiments of the present disclosure, the liquidpharmaceutical formulations may include leucine (e.g., L-leucine) at aconcentration of from about 1 mM to about 250 mM±10%. In someembodiments of the present disclosure, the liquid pharmaceuticalformulations may include leucine at a concentration of about 100 mM±10%or at a concentration of 150 mM±10%.

In certain embodiments of the present disclosure, the liquidpharmaceutical formulations may include methionine (e.g., L-methionine)at a concentration of about 1 mM to about 100 mM±10%. In someembodiments of the present disclosure, the liquid pharmaceuticalformulations may include methionine at a concentration of about 5 mM toabout 15 mM±10%. In other embodiments of the present disclosure, theliquid pharmaceutical formulations may include methionine at aconcentration of about 10 mM±10%.

In certain embodiments of the present disclosure, the liquidpharmaceutical formulations may include proline (e.g., L-proline) at aconcentration of from about 5 mM to about 300 mM±10%. In otherembodiments of the present disclosure, the liquid pharmaceuticalformulations may include proline at a concentration of from about 25 toabout 75 mM±10%. In other embodiments of the present disclosure, theliquid pharmaceutical formulations may include proline at aconcentration of about 50 mM±10%.

In a further aspect provided the pharmaceutical formulation may furtherinclude a surfactant.

Exemplary embodiments of surfactants include, for example and withoutlimitations, non-ionic surfactants. Surfactant described herein include,for example, polysorbate 20 (PS-20), polysorbate 40 (PS-40), polysorbate60 (PS-60), polysorbate 65 (PS-65), polysorbate 80 (PS-80), TritonX-100, poloxamer, Pluronic F-68 and the like including combinationsthereof.

The concentration of the surfactant may be from about 0.001 to about0.1%; from about 0.005 to about 0.05%; from about 0.01 to about 0.05%;from about 0.01 to about 0.04%; from about 0.01 to about 0.03%; fromabout 0.015 to about 0.025% etc. The concentration of the surfactant maybe for example, about 0.02%±10%. However, it is to be understood thatthe concentration of the surfactant can be either higher or lower asspecific conditions require. The concentration of surfactant isgenerally expressed as weight/volume (w/v). However, in some instances,the concentration of surfactant may be expressed as volume/volume (v/v).

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, polysorbate 20 at a concentration in the rangeof from about 0.001 to about 0.1% (w/v), from about 0.005 to about0.05%, from (w/v) about 0.01 to about 0.05%, from about 0.01 to about0.03% (w/v) etc. In an exemplary embodiment, polysorbate 20 may be at aconcentration about 0.02 (w/v)±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, polysorbate 40 at a concentration in the rangeof from about 0.001 to about 0.1% (w/v), from about 0.005 to about 0.05%(w/v), from about 0.01 to about 0.05% (w/v), from about 0.01 to about0.03% (w/v) etc. In an exemplary embodiment, polysorbate 40 may be at aconcentration about 0.02 (w/v)±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, polysorbate 60 at a concentration in the rangeof from about 0.001 to about 0.1% (w/v), from about 0.005 to about 0.05%(w/v), from about 0.01 to about 0.05% (w/v), from about 0.01 to about0.03% (w/v) etc. In an exemplary embodiment, polysorbate 60 may be at aconcentration about 0.02 (w/v)±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, polysorbate 80 at a concentration in the rangeof from about 0.001 to about 0.1% (w/v), from about 0.005 to about 0.05%(w/v), from about 0.01 to about 0.05% (w/v), from about 0.01 to about0.03% (w/v) etc. In an exemplary embodiment, polysorbate 80 may be at aconcentration about 0.02 (w/v)±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, Triton X-100 at a concentration in the rangeof from about 0.001 to about 0.1%, from about 0.005 to about 0.05%, fromabout 0.01 to about 0.05%, from about 0.01 to about 0.03% etc. In anexemplary embodiment, Triton X-100 may be at a concentration about 0.02(v/v)±10%.

In some embodiments described herein, the pharmaceutical formulationsmay include, for example, Pluronic F-68 at a concentration in the rangeof from about 0.001 to about 0.1%, from about 0.005 to about 0.05%, fromabout 0.01 to about 0.05%, from about 0.01 to about 0.03% etc. In anexemplary embodiment, Pluronic F-68 may be at a concentration about 0.02(v/v)±10%.

In certain aspects and embodiments of the present disclosure, thepharmaceutical formulations described herein may have a pH in the rangeof from about 5.0 to about 8.0, from about 5.0 to about 7.5; from about5.5 to about 7.5; from about 6.0 to about 8.0; from about 6.0 to about7.5; from about 6.0 to about 7.0; from about 6.5 to about 7.5; fromabout 7.0 to about 7.5 etc. The pH of the pharmaceutical formulationsmay be for example about, 6.0±0.15, 6.1±0.15, 6.2±0.15, 6.3±0.15,6.4±0.15, 6.5±0.15, 6.6±0.15, 6.7±0.15, 6.8±0.15, 6.9±0.15, 7.0±0.15,7.1±0.15, 7.2±0.15, 7.3±0.15, 7.4±0.15 or 7.5±0.15.

In an exemplary embodiment, the pH of the pharmaceutical formulationsmay be for example about 6.5±0.5.

In an exemplary embodiment, the pH of the pharmaceutical formulationsmay be for example about 7.0±0.5.

In an exemplary embodiment, the pH of the pharmaceutical formulationsmay be for example about 7.5±0.5.

The pharmaceutical formulation of the present disclosure may have a pHof between 6.0±0.15 to 7.5±0.15. Alternatively, the pharmaceuticalformulation of the present disclosure may have a pH of between 6.5±0.15to 7.0±0.15. Exemplary embodiments disclosed herein include,pharmaceutical formulation having a pH of 6.0±0.15. Other exemplaryembodiments disclosed herein include, pharmaceutical formulation havinga pH of 6.5±0.15. Further exemplary embodiments disclosed hereininclude, pharmaceutical formulation having a pH of 7.0±0.15. Furtherexemplary embodiments disclosed herein include, pharmaceuticalformulation having a pH of 7.5±0.15.

In exemplary embodiments, the pharmaceutical formulations describedherein may have a pH of approximately 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6,6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8.0at room temperature (e.g., approximately 23° C.).

In other exemplary embodiments, the pharmaceutical formulationsdescribed herein may have a pH of approximately 6.3, 6.4, 6.5, 6.6, 6.7,6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7 at room temperature(e.g., approximately 23° C.).

In yet other exemplary embodiments, the pharmaceutical formulationsdescribed herein may have a pH of approximately 6.5, 6.6, 6.7, 6.8, 6.9,7.0, 7.1, 7.2, 7.3, 7.4, 7.5 at room temperature (e.g., approximately23° C.).

The liquid pharmaceutical formulations of the present disclosure may bestable under stress conditions. For example, the liquid formulation maybe stable under mechanical stress and/or under freeze-thawingconditions.

Exemplary embodiments and aspects of the present disclosure relate topharmaceutical formulations that may be stable for at least 1 monthunder refrigerated conditions (e.g., 2-8° C. including 3° C., 4° C., 5°C., 6° C., 7° C.). Exemplary and aspects of the present disclosurerelate to pharmaceutical formulations that may be stable for at least 2months; for at least 3 months or for at least 6 months underrefrigerated conditions. Exemplary and aspects of the present disclosurealso relate to pharmaceutical formulations that may be stable for atleast 12 months under refrigerated conditions. Exemplary and aspects ofthe present disclosure also relate to pharmaceutical formulations thatmay be stable for at least 18 months under refrigerated conditions.

Further exemplary embodiments and aspects of the present disclosurerelate to pharmaceutical formulations that may be stable for at least 1month, for at least 2 months, for at least 3 months at room temperature(e.g., 15° C.-30° C. including at 25° C.). Additional exemplaryembodiments and aspects of the present disclosure relate topharmaceutical formulations that may be stable for at least 6 months atroom temperature. Other exemplary embodiments and aspects of the presentdisclosure relate to pharmaceutical formulations that may be stable forat least 12 months at room temperature.

Moreover, additional exemplary embodiments and aspects of the presentdisclosure relate to pharmaceutical formulations that may be stable forat least 1 months, for at least 2 months, or for at least 3 months atelevated temperature (at least 40° C.).

In a further aspect provided is a liquid pharmaceutical formulation thatincludes a glucagon-like peptide 1 (GLP-1) or a GLP-1 analogue, abuffering agent, a tonicity agent, and optionally an excipient and/or asurfactant.

In yet a further aspect provided is a liquid pharmaceutical formulationthat includes a glucagon-like peptide 1 (GLP-1) or a GLP-1 analogue, abuffering agent, a tonicity agent, an excipient and optionally asurfactant.

In yet a further aspect provided is a liquid pharmaceutical formulationthat includes a glucagon-like peptide 1 (GLP-1) or a GLP-1 analogue, abuffering agent, a tonicity agent, a surfactant and optionally anexcipient.

In yet additional aspects provided a liquid pharmaceutical formulationthat includes a glucagon-like peptide 1 (GLP-1) or a GLP-1 analogue, abuffering agent, a tonicity agent, an excipient and a surfactant.

Numerous and non-limiting embodiments of pharmaceutical formulations ofthe present disclosure are provided below.

In some embodiments, the pharmaceutical formulation is at a pH of about6.0±0.5 to 7.5±0.5 and includes a GLP-1 or a GLP-1 analogue, a bufferingagent, and a tonicity agent at a concentration sufficient to allowisotonicity with blood and/or serum.

In some embodiments, the pharmaceutical formulation is at a pH of about6.0±0.15 to 7.5±0.15 and includes a GLP-1 or a GLP-1 analogue, abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

In additional embodiments, the pharmaceutical formulation is at a pH ofabout 6.0±0.5 to 7.5±0.5 and includes a GLP-1 analogue comprising orconsisting of the amino acid sequence set forth in SEQ ID NO:3; abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

In additional embodiments, the pharmaceutical formulation is at a pH ofabout 6.0±0.15 to 7.5±0.15 and includes a GLP-1 analogue comprising orconsisting of the amino acid sequence set forth in SEQ ID NO:3; abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

In additional embodiments, the pharmaceutical formulation is at a pH ofabout 6.0±0.15 to 7.5±0.5 and includes a GLP-1 analogue comprising orconsisting of the amino acid sequence set forth in SEQ ID NO:3; abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

In further embodiments, the pharmaceutical formulation is at a pH ofabout 6.5±0.15 to 7.0±0.15 and includes a GLP-1 analogue comprising orconsisting of the amino acid sequence set forth in SEQ ID NO:3, abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

In further embodiments, the pharmaceutical formulation is at a pH ofabout 6.5±0.5 to 7.0±0.5 and includes a GLP-1 analogue comprising orconsisting of the amino acid sequence set forth in SEQ ID NO:3, abuffering agent, and a tonicity agent at a concentration sufficient toallow isotonicity with blood and/or serum.

The pharmaceutical formulation may further comprise an excipient and/orsurfactant. The surfactant may be non-ionic and may include for example,polysorbate (e.g., PS-20, PS-40, PS-60, PS-80 and the like). Theexcipient may be an amino acid excipient.

In aspects and embodiments described herein, the pharmaceuticalformulation comprises a GLP-1 analogue having the amino acid sequenceset forth in SEQ ID NO:3, wherein the liquid pharmaceutical formulationis at a pH of between 6.5±0.15 and 7.0±0.15 and comprises histidine-HClas buffering agent, sucrose or mannitol as tonicity agent, methionine orproline as an excipient and a surfactant.

In some embodiments, the liquid pharmaceutical formulation comprises aGLP-1 analogue at a concentration of from about 50 μg/ml to about 2mg/ml.

In some embodiments, the liquid pharmaceutical formulation comprises aGLP-1 analogue at a concentration of from about 50 μg/ml to about 1mg/ml.

In some embodiments, the liquid pharmaceutical formulation comprises aGLP-1 analogue at a concentration of from about 100 μg/ml to about 1mg/ml.

In some embodiments, the liquid pharmaceutical formulation comprises aGLP-1 analogue at a concentration of from about 100 μg/ml to about 500μg/ml.

In some embodiments, the surfactant is polysorbate-20.

In some embodiments, the liquid pharmaceutical formulation comprises aGLP-1 analogue at a concentration of from about 300 μg/ml±20%.

In some embodiments, the liquid pharmaceutical formulation is at a pH of6.5±0.5.

In some embodiments, the liquid pharmaceutical formulation is at a pH of7.0±0.5.

In some embodiments, the liquid pharmaceutical formulation is at a pH of7.0±0.15.

In some embodiments, the liquid pharmaceutical formulation is at a pH of7.5±0.5.

In aspects and embodiments described herein, the pharmaceuticalformulation is at a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to7.5±0.15 and includes a) a GLP-1 analogue comprising or consisting ofthe amino acid sequence set forth in SEQ ID NO:3 at a concentration offrom about 10 μg/ml to about 1 mg/ml, b) a citrate buffer at aconcentration of from about 1 mM to about 50 mM, c) glycerol at aconcentration of from about 10 mM to about 500 mM, mannitol at aconcentration of from about 10 mM to about 500 mM, sucrose at aconcentration of from about 1% to about 20%, trehalose at aconcentration of from about 1% to about 20% or NaCl at a concentrationof from about 50 mM to about 150 mM, d) optionally polysorbate at aconcentration of from about 0.001 to about 0.1% (w/v), and e) optionallyan amino acid excipient at a concentration of about 0.5 mM to about 300mM.

In aspects and embodiments described herein, the pharmaceuticalformulation is at a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to7.5±0.15 and includes a) a GLP-1 analogue comprising or consisting ofthe amino acid sequence set forth in SEQ ID NO:3 at a concentration offrom about 10 μg/ml to about 1 mg/ml, b) a histidine buffer at aconcentration of from about 1 mM to about 50 mM, c) glycerol at aconcentration of from about 10 mM to about 500 mM, mannitol at aconcentration of from about 10 mM to about 500 mM, sucrose at aconcentration of from about 1% to about 20%, trehalose at aconcentration of from about 1% to about 20% or NaCl at a concentrationof from about 50 mM to about 150 mM, d) optionally polysorbate at aconcentration of from about 0.001 to about 0.1% (w/v), and e) optionallyan amino acid excipient at a concentration of about 0.5 mM to about 300mM.

In further aspect and embodiments described herein, the pharmaceuticalformulation is at a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to7.5±0.15 and includes a) a GLP-1 analogue comprising or consisting ofthe amino acid sequence set forth in SEQ ID NO:3 at a concentration offrom about 10 μg/ml to about 1 mg/ml, b) a phosphate buffer at aconcentration of from about 1 mM to about 50 mM, c) glycerol at aconcentration of from about 10 mM to about 500 mM, mannitol at aconcentration of from about 10 mM to about 500 mM, sucrose at aconcentration of from about 1% to about 20%, trehalose at aconcentration of from about 1% to about 20% or NaCl at a concentrationof from about 50 mM to about 150 mM, d) optionally polysorbate at aconcentration of from about 0.001 to about 0.1% (w/v), and e) optionallyan amino acid excipient at a concentration of about 0.5 mM to about 300mM.

In further aspect and embodiments described herein, the pharmaceuticalformulation is at a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to7.5±0.15 and includes a) a GLP-1 analogue comprising or consisting ofthe amino acid sequence set forth in SEQ ID NO:3 at a concentration offrom about 10 μg/ml to about 1 mg/ml, b) a maleate buffer at aconcentration of from about 1 mM to about 50 mM, c) glycerol at aconcentration of from about 10 mM to about 500 mM, mannitol at aconcentration of from about 10 mM to about 500 mM, sucrose at aconcentration of from about 1% to about 20%, trehalose at aconcentration of from about 1% to about 20% or NaCl at a concentrationof from about 50 mM to about 150 mM, d) optionally polysorbate at aconcentration of from about 0.001 to about 0.1% (w/v), and e) optionallyan amino acid excipient at a concentration of about 0.5 mM to about 300mM.

In additional aspects and embodiments, the pharmaceutical formulation isat a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to 7.5±0.15 andincludes a GLP-1 or a GLP-1 analogue comprising or consisting of theamino acid sequence set forth in SEQ ID NO:3 at a concentration of fromabout 100 μg/ml to about 500 μg/ml, citrate at a concentration of fromabout 7.5 mM to about 12.5 mM±10% and glycerol at a concentration offrom about 150 mM to about 300 mM±10%, mannitol at a concentration offrom about 150 mM to about 300 mM±10%, sucrose at a concentration offrom about 5% to about 15%±10% or trehalose at a concentration of fromabout 5% to about 15%±10%.

In additional aspects and embodiments, the pharmaceutical formulation isat a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to 7.5±0.15 andincludes a GLP-1 or a GLP-1 analogue comprising or consisting of theamino acid sequence set forth in SEQ ID NO:3 at a concentration of fromabout 100 μg/ml to about 500 μg/ml, histidine-HCl at a concentration offrom about 7.5 mM to about 12.5 mM±10% and glycerol at a concentrationof from about 150 mM to about 300 mM±10%. mannitol at a concentration offrom about 150 mM to about 300 mM±10%, sucrose at a concentration offrom about 5% to about 15%±10% or trehalose at a concentration of fromabout 5% to about 15%±10%.

In other aspects and embodiments, the pharmaceutical formulation is at apH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to 7.5±0.15 and includes aGLP-1 or a GLP-1 analogue comprising or consisting of the amino acidsequence set forth in SEQ ID NO:3 at a concentration of from about 100μg/ml to about 500 μg/ml, sodium phosphate at a concentration of fromabout 7.5 mM to about 12.5 mM±10% and glycerol at a concentration offrom about 150 mM to about 300 mM±10%, mannitol at a concentration offrom about 150 mM to about 300 mM±10%, sucrose at a concentration offrom about 5% to about 15%±10% or trehalose at a concentration of fromabout 5% to about 15%±10%.

In other aspects and embodiments, the pharmaceutical formulation is at apH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to 7.5±0.15 and includes aGLP-1 or a GLP-1 analogue comprising or consisting of the amino acidsequence set forth in SEQ ID NO:3 at a concentration of from about 100μg/ml to about 500 μg/ml, potassium phosphate at a concentration of fromabout 7.5 mM to about 12.5 mM±10% and glycerol at a concentration offrom about 150 mM to about 300 mM±10%, mannitol at a concentration offrom about 150 mM to about 300 mM±10%, sucrose at a concentration offrom about 5% to about 15%±10% or trehalose at a concentration of fromabout 5% to about 15%±10%.

In additional aspects and embodiments, the pharmaceutical formulation isat a pH of about 6.5±0.5 to 7.5±0.5, or 6.0±0.15 to 7.5±0.15 andincludes a GLP-1 or a GLP-1 analogue comprising or consisting of theamino acid sequence set forth in SEQ ID NO:3 at a concentration of fromabout 100 μg/ml to about 500 μg/ml, maleate at a concentration of fromabout 7.5 mM to about 12.5 mM±10% and glycerol at a concentration offrom about 150 mM to about 300 mM±10%, mannitol at a concentration offrom about 150 mM to about 300 mM±10%, sucrose at a concentration offrom about 5% to about 15%±10% or trehalose at a concentration of fromabout 150 mM to about 300 mM±10%.

In additional embodiments, the pharmaceutical formulation furthercomprises a surfactant and/or an amino acid excipient. For example, thepharmaceutical formulations may comprise polysorbate at a concentrationof from about 0.005 to about 0.05% (w/v)±10% and/or an amino acidexcipient.

In further embodiment, the pharmaceutical formulation comprises an aminoacid excipient such as methionine at a concentration of from about 5 mMto about 25 mM±10%, proline at a concentration of from about 25 mM toabout 75 mM±10%, glycine at a concentration of from about 50 mM to about150 mM±10% and/or arginine at a concentration of from about 50 mM toabout 150 mM±10%.

In some embodiments, pharmaceutical formulation comprising histidinebuffer may be at pH of about 6.5±0.5 to 7.5±0.5. In other embodiments,pharmaceutical formulation comprising phosphate buffer may be at pH ofabout 6.5±0.5 to 7.5±0.5. In other embodiments, the pharmaceuticalformulation is at pH of about 6.5±0.15 to 7.0±0.15. In additionalembodiments, pharmaceutical formulations that comprise citrate bufferare at a pH of about 6.5±0.15. In other embodiments, pharmaceuticalformulations that comprise histidine buffer are at a pH of about7.0±0.15. In additional embodiments, pharmaceutical formulations thatcomprise maleate buffer are at a pH of about 7.0±0.15. In furtherembodiments, pharmaceutical formulations that comprise phosphate bufferare at a pH of about 6.5±0.15.

In additional embodiments, the concentration of the GLP-1 analogue setforth in SEQ ID NO:3 in the pharmaceutical formulation is from about 10μg/ml to about 1 mg/ml. In further embodiments, the GLP-1 analogue setforth in SEQ ID NO:3 is at a concentration of from about 100 μg/ml toabout 500 μg/ml. In yet further embodiments the GLP-1 analogue set forthin SEQ ID NO:3 is at a concentration of about 0.3 mg/ml±20%. In otherembodiments the GLP-1 analogue set forth in SEQ ID NO:3 is at aconcentration of about 0.3 mg/ml±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15, etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, sodium citrate at a concentration of about 10 mM±10%, atonicity agent such as glycerol at a concentration of from about 165 mMto about 275 mM±10%, sucrose at a concentration of from about 5% toabout 15%, trehalose at a concentration of from about 5% to about 15% orNaCl at a concentration of from about 50 mM to about 150 mM, asurfactant such as polysorbate at a concentration of about 0.005 to0.05% (w/v)±10% and an amino acid excipient such as methionine at aconcentration of about 5 mM to about 25 mM±10%, proline at aconcentration of from about 25 mM to about 75 mM±10% or arginine at aconcentration of from about 50 mM to about 150 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15, etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, histidine HCl at a concentration of about 10 mM±10%, atonicity agent such as glycerol at a concentration of from about 165 mMto about 275 mM±10%, sucrose at a concentration of from about 5% toabout 15%, trehalose at a concentration of from about 5% to about 15% orNaCl at a concentration of from about 50 mM to about 150 mM, asurfactant such as polysorbate at a concentration of about 0.005 to0.05% (w/v)±10% and an amino acid excipient such as methionine at aconcentration of about 5 mM to about 25 mM±10%, proline at aconcentration of from about 25 mM to about 75 mM±10% or arginine at aconcentration of from about 50 mM to about 150 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15, etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, histidine HCl at a concentration of about 10 mM±10%, atonicity agent such as mannitol at a concentration of from about 165mM±10% to about 275 mM±10%, or sucrose at a concentration of from about5%±10% to about 15%±10%, a surfactant such as polysorbate-20 at aconcentration of about 0.005±10% to 0.05%±10% (w/v)±10% and an aminoacid excipient such as methionine at a concentration of about 5 mM±10%to about 25 mM±10%, or proline at a concentration of from about 25mM±10% to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15 etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, maelate at a concentration of about 10 mM±10%, a tonicityagent such as glycerol at a concentration of from about 165 mM to about275 mM±10%, sucrose at a concentration of from about 5% to about 15%,trehalose at a concentration of from about 5% to about 15% or NaCl at aconcentration of from about 50 mM to about 150 mM, a surfactant such aspolysorbate at a concentration of about 0.005 to 0.05% (w/v)±10% and anamino acid excipient such as methionine at a concentration of about 5 mMto about 25 mM±10%, proline at a concentration of from about 25 mM toabout 75 mM±10% or arginine at a concentration of from about 50 mM toabout 150 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15, etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, sodium phosphate at a concentration of about 10 mM±10%, atonicity agent such as glycerol at a concentration of from about 165 mMto about 275 mM±10%, sucrose at a concentration of from about 5% toabout 15%, trehalose at a concentration of from about 5% to about 15% orNaCl at a concentration of from about 50 mM to about 150 mM, asurfactant such as polysorbate at a concentration of about 0.005 to0.05% (w/v)±10% and an amino acid excipient such as methionine at aconcentration of about 5 mM to about 25 mM±10%, proline at aconcentration of from about 25 mM to about 75 mM±10% or arginine at aconcentration of from about 50 mM to about 150 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 6.5±0.15 to 7.0±0.15, etc.) and includes aGLP-1 analogue comprising or consisting of the amino acid sequence setforth in SEQ ID NO:3 at a concentration of from about 100 μg/ml to about500 μg/ml, potassium phosphate at a concentration of about 10 mM±10%, atonicity agent such as glycerol at a concentration of from about 165 mMto about 275 mM±10%, sucrose at a concentration of from about 5% toabout 15%, trehalose at a concentration of from about 5% to about 15% orNaCl at a concentration of from about 50 mM to about 150 mM, asurfactant such as polysorbate at a concentration of about 0.005 to0.05% (w/v)±10% and an amino acid excipient such as methionine at aconcentration of about 5 mM to about 25 mM±10%, proline at aconcentration of from about 25 mM to about 75 mM±10% or arginine at aconcentration of from about 50 mM to about 150 mM±10%.

Exemplary embodiments include pharmaceutical formulations at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15 etc.) and including a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, and mannitolat a concentration of about 165 mM to about 275 mM±10%, sucrose at aconcentration of about 5% to about 15%±10% or trehalose at aconcentration of about 5% to about 15%±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15 etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 165 mM to about 275 mM±10%, and polysorbate-20at a concentration of about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 275 mM±10% and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 165 mM±10% and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 275 mM±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and methionine at a concentration ofabout 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 165 mM±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and methionine at a concentration ofabout 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 275 mM±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and proline at a concentration of about25 mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, mannitol ata concentration of about 165 mM±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and proline at a concentration of about25 mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 5% to about 15%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 8%±10%, and polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 10%±10%, and polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 10%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 10%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 5% to about 15%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 8%±10%, and polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 10%±10%, and polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 10%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, histidine-HCl at a concentration of about 10 mM±10%, trehalose ata concentration of about 10%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, andmannitol at a concentration of about 165 mM to about 275 mM±10%, sucroseat a concentration of about 5% to about 15%±10% or trehalose at aconcentration of about 5% to about 15%±10.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 165 mM to about 275 mM±10%, andpolysorbate-20 at a concentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 275 mM±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 165 mM±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 275 mM±10%, polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10% and methionine at aconcentration of about 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 165 mM±10%, polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10% and methionine at aconcentration of about 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 275 mM±10%, polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10% and proline at aconcentration of about 25 mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, mannitolat a concentration of about 165 mM±10%, polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10% and proline at aconcentration of about 25 mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 5% to about 15%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 8%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 10%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 10%±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and methionine at a concentration ofabout 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, sucroseat a concentration of about 10%±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and proline at a concentration of about25 mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 5% to about 15%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 8%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In other embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 10%±10%, and polysorbate-20 at aconcentration of about 0.01 to 0.03% (w/v)±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and methionine at a concentration of about5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 10%±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and methionine at a concentration ofabout 5 mM to about 25 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 8%±10%, polysorbate-20 at a concentration ofabout 0.01 to 0.03% (w/v)±10% and proline at a concentration of about 25mM to about 75 mM±10%.

In further embodiments, the pharmaceutical formulation is at a pH of6.5±0.5 to 7.5±(e.g., 7.0±0.5, 6.5±0.15, etc.) and includes a GLP-1analogue comprising or consisting of the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from about 100 μg/ml to about 500μg/ml, sodium phosphate at a concentration of about 10 mM±10%, trehaloseat a concentration of about 10%±10%, polysorbate-20 at a concentrationof about 0.01 to 0.03% (w/v)±10% and proline at a concentration of about25 mM to about 75 mM±10%.

The pharmaceutical formulation of the present disclosure may be used forthe treatment of irritable bowel syndrome (IBS). In exemplaryembodiments, the pharmaceutical formulation may be used for relief ofpain in IBS and/or for relief of constipation in IBS-C. Thepharmaceutical formulation may be used for the treatment of IBS-C, ofdiarrhea predominant IBS (IBS-D) or of mixed or alternating type IBS(IBS-M). Prevention of pain in IBS, IBS-C, IBS-D and/or IBS-M is alsocontemplated.

Alternatively, the formulation of the present disclosure may be used fortreatment of other diseases or conditions including without limitationdiabetes, ischemia, reperfused tissue injury, dyslipidemia, diabeticcardiomyopathy, myocardial infarction, acute coronary syndrome, obesity,catabolic changes after surgery, hyperglycemia, irritable bowelsyndrome, stroke, neurodegenerative disorders, memory and learningdisorders, islet cell transplant, functional dyspepsia and/orregenerative therapy in mammals in need of treatment.

The term “treatment” for purposes of this disclosure refers to boththerapeutic treatment and prophylactic or preventative measures. Thosein need of treatment include those already with the disorder as well asthose prone to have the disorder or those in whom the disorder is to beprevented.

In certain aspects and embodiments, the present disclosure thereforeprovides a stable liquid formulation at a pH of between 6.0±0.15 to7.5±0.15 and that includes glucagon-like peptide 1 (GLP-1) or a GLP-1analogue thereof, a buffering agent, and optionally a tonicity agent,excipient and/or surfactant, wherein the formulation is for use in themaking of a stable pharmaceutical formulation. For example, a stablepharmaceutical formulation can be obtained by adding a tonicity agent asdescribed herein and/or excipient as described herein to such stableliquid formulation.

In other aspects and embodiments, the present disclosure provides amethod of making a stable pharmaceutical formulation that include addingan excipient and/or a surfactant to a stable formulation so as tofurther stabilize it,

Further scope, applicability and advantages will become apparent fromthe non-restrictive detailed description given hereinafter. It should beunderstood, however, that this detailed description, while indicatingexemplary embodiments or aspects, is given by way of example only.

DETAILED DESCRIPTION

The use of the terms “a” and “an” and “the” are to be construed to coverboth the singular and the plural, unless otherwise indicated herein orclearly contradicted by context.

Unless specifically stated or obvious from context, as used herein theterm “or” is understood to be inclusive and covers both “or” and “and”.

The term “and/or” where used herein is to be taken as specificdisclosure of each of the specified features or components with orwithout the other.

The terms “comprising”, “having”, “including”, and “containing” are tobe construed as open-ended terms (i.e., meaning “including, but notlimited to”) unless otherwise noted. The term “consisting of” is to beconstrued as close-ended.

The term “about” or “approximately” shall generally mean within 20percent, within 10 percent, within 5, 4, 3, 2 or 1 percent of a givenvalue or range.

As used herein the term “liquid” with respect to formulations means thatthe peptide is dissolved in a solution without significantprecipitation.

As used herein the term “stable” with respect to a formulation refers toa formulation in which no significant aggregation and/or degradation ofthe GLP-1 or GLP-1 analogue occurs either in absence of stress or underone or more stress conditions.

As used herein the term “no significant aggregation and/or degradation”means aggregation and/or degradation of less than 20 percent, of lessthan 15 percent, less than 10 percent, less than 5 percent, less than 3percent or less than 2 percent. For example, a formulation that showsless than 5% degradation after 1-3 months storage at 5° C., less than 10percent degradation after 3 months storage at 25° C. and/or less than 20percent degradation after 3 months storage at 40° C. may be consideredstable. A formulation that shows less than 10 percent or even less than5 percent degradation after freeze-thaw and/or mechanical stress may beconsidered stable.

As used herein the term “one or more stress conditions” refers toprolonged storage (e.g., under refrigerated conditions), acceleratedstorage conditions (e.g., ambient temperature and/or elevatedtemperature such as for example 40° C.), mechanical stress,freeze-thawing or combination thereof.

As used herein the term “high concentration of subvisible particles”encompasses for example a cumulative particle count of more than 1,000for particles of ≥2 μm, more than 500 for particles of ≥5 μm, more than100 particles of ≥10 μm and/or of more than 10 particles of ≥25 μm asmeasured by micro-flow imaging at either T0 in unstressed formulations,T−1m_25° C. and/or T−1m_40° C.

As used herein the term “low concentration of subvisible particles”encompasses for example cumulative particle counts of 1000 or below forparticles ≥2 μm, 275 or below for particles ≥5 μm, 100 or below forparticles ≥10 μm and/or 30 or below for particles ≥25 μm as measured bymicro-flow imaging at T−1m_25° C. and/or T−1m_40° C. The term “lowconcentration of subvisible particles” also encompasses cumulativeparticle counts of 500 or below for particles ≥2 μm, 200 or below forparticles ≥2 μm, 50 or below for particles ≥10 μm and 10 and/or belowfor particles 25 μm as measured by micro-flow imaging at T−3m_5° C.and/or T−3m_25° C.

The term “sugar” refers to monosaccharides, disachharides, andpolysaccharides. Examples of sugars include, but are not limited to,sucrose, glucose, dextrose, and others.

The term “long-term storage” or “prolonged storage” is understood tomean storage of at least one month, at least three months, at least sixmonths and/or at least one year.

The term “mammal” includes, but is not limited to, a human.

The terms “pharmaceutical composition” and “pharmaceutical formulation”are used interchangeably.

Liquid pharmaceutical formulations of GLP-1 or of GLP-1 analogue thereofare provided. The liquid pharmaceutical formulations of the presentdisclosure support physical stress such as freeze-thaw, mechanicalstress, elevated temperature and/or long-term storage.

In some instances, liquid pharmaceutical formulations that include theGLP-1 or of GLP-1 analogue, a buffering agent and a tonicity agent havebeen found to be stable without excipient or surfactant. Liquidpharmaceutical formulations that include GLP-1 or the GLP-1 analoguethereof a buffering agent, a tonicity agent, and optionally an excipientand/or surfactant are therefore provided.

In other instances, the present disclosure relates to liquidpharmaceutical formulations that include the GLP-1 or of GLP-1 analogue,a buffering agent, a tonicity agent, a surfactant and an excipient.

Embodiments disclosed herein relate to liquid pharmaceuticalformulations that include ROSE-010 (Val8-GLP-1). The formulations of thepresent disclosure can comprise ROSE-010 peptide concentration of fromabout 10 μg/ml to about 2 mg/ml, from about 50 μg/ml to about 2 mg/ml,from about 100 μg/ml to about 2 mg/ml, from about 10 μg/ml to about 1mg/ml, from about 100 μg/ml to about 1 mg/ml, from about 50 μg/ml toabout 750 μg/ml, from about 100 μg/ml to about 500 μg/ml, from about 200μg/ml to about 500 μg/ml, at least 1 mg/ml, at least 750 μg/ml, at least500 μg/ml, at least 300 μg/ml and including 300 μg/ml±20% or 300μg/ml±10%.

The pharmaceutical formulations appear more stable at a higher pH (e.g.,pH 6.5±0.5 to 7.5±0.5, pH 6.0±0.15 to 7.5±0.15). For example, a pH of6.5±0.5 to 7.5±0.5 (e.g., including 6.5±0.15 to 7.0±0.15) may bebeneficial for peptide stability. In exemplary embodiments, a pH of6.5±0.5, 7.0±0.5 (e.g., 7.0±0.15), or 7.5±0.5 may be provided byhistidine buffer. In other exemplary embodiments, a pH of 6.5±0.5,(e.g., 6.5±0.15), 7.0±0.5, or 7.5±0.5 may be provided by a sodiumphosphate buffer or potassium phosphate buffer.

Stability of the formulations does not appear to be influenced by thetonicity agents. However, pharmaceutical formulations that containtonicity agents are encompassed by the present disclosure. The tonicityagent can be sugars or simpler and less reactive tonicity agents.However, other tonicity agents can be used. The formulations may berendered more stable by the addition of a surfactant and/or excipient orboth a surfactant and an excipient.

Surfactants may thus be beneficial for the stability of thepharmaceutical formulations disclosed herein. Non-ionic surfactants suchas polysorbates (e.g., PS-20, PS-40, PS-60, PS-65, PS-80 etc.), TritonX-100, poloxamer, Pluronic F-68 and the like and combination thereof canbe added to the pharmaceutical formulations so as to protect GLP-1 orthe GLP-1 analogues from degradation and/or aggregation.

Pharmaceutical formulations that contain polysorbates appear to protectGLP-1 or the GLP-1 analogues during mechanical stress. Polysorbates maythus help to stabilize ROSE-010 against mechanical stress, such asduring shipment and product handling.

Pharmaceutical formulations that contain small amounts of amino acidexcipients appear more stable.

Exemplary embodiments of pharmaceutical formulations that are disclosedherein are stable under common stress conditions, such as freeze-thawstress and/or mechanical stress. The purity of exemplary pharmaceuticalformulations disclosed herein judged by RP-UPLC is about 95% afterstorage at 25° C. for up to at least 3 months. Exemplary pharmaceuticalformulations disclosed herein appear stable under storage temperature of2-8° C. (e.g., for up to at least three months).

The liquid formulation of the present disclosure may be suitable forinjection. In certain aspects and embodiments described herein theliquid formulation may be provided in pre-filled syringes. In otheraspects and embodiments described herein, the liquid formulation may beprovided in single-dose or multiple-dose containers. In yet otheraspects and embodiments described herein the liquid formulation maysustain prolonged storage (long-term storage).

In exemplary embodiments, the single-dose or multiple-dose containerinclude for example, vials (e.g. glass vials), syringes (e.g.,pre-filled syringes), cartridges, injectors (e.g., pen system and othertypes of auto-injectors) and the like.

The formulation may also be suitable for use in non-parenteral delivery.In certain aspects, the formulation may be encapsulated for oral ormucosal (e.g., nasal) delivery.

The formulation of the present disclosure may contain doses of GLP-1 orGLP-1 analogues of at least 20 μg, at least 30 μg, at least 50 μg, atleast 75 μg, at least 100 μg, at least 150 μg, at least 200 μg, at least300 μg, at least 500 μg, at least 1 mg. The dose contained in each vialor syringe will depend on the need of the patient population.

Since liquid formulations that contains the GLP-1 or of GLP-1 analogue,a buffering agent and a tonicity agent have been found to be stablewithout excipient or surfactant, a liquid formulation (e.g. stocksolution) that contains these elements may be prepared and stored for atleast one week (i.e. at least two weeks, at least three weeks, at leastone month, at least two months, at least three months etc.). Apharmaceutical composition may subsequently be reconstituted by addingother elements to make it suitable for longer storage.

In addition to the embodiments described and provided in thisdisclosure, the following non-limiting embodiments are particularlycontemplated.

-   -   1—A pharmaceutical formulation including a GLP-1 or a GLP-1        analogue, a buffering agent, a tonicity agent and optionally an        excipient and/or surfactant.    -   2—A pharmaceutical formulation including a GLP-1 or a GLP-1        analogue, a buffering agent, a tonicity agent, an excipient        and/or a non-ionic surfactant.    -   3—A pharmaceutical formulation as individually or collectively        listed in Table 1A and including a GLP-1 or a GLP-1 analogue.    -   4—A pharmaceutical formulation as individually or collectively        listed in Table 1B and including a GLP-1 or a GLP-1 analogue.    -   5—A pharmaceutical formulation as individually or collectively        listed in Table 10 and including a GLP-1 or a GLP-1 analogue.    -   6—A pharmaceutical formulation as individually or collectively        listed in Table 1D and including a GLP-1 or a GLP-1 analogue.    -   7—A pharmaceutical formulation as individually or collectively        listed in Table 1E and including a GLP-1 or a GLP-1 analogue.    -   8—A pharmaceutical formulation as individually or collectively        listed in Table 4 and including a GLP-1 or a GLP-1 analogue.    -   9—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is GLP-1        (7-37) and comprises the amino acid sequence set forth in SEQ ID        NO:2.    -   10—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is ROSE-010        and comprises the amino acid sequence set forth in SEQ ID NO:3.    -   11—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 analogue comprises an amino acid        sequence at least 90% identical to SEQ ID NO:3 wherein the amino        acid at position 2 is valine.    -   12—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 50 μg to the user.    -   13—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 100 μg to the user.    -   14—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 150 μg to the user.    -   15—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 200 μg to the user.    -   16—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 300 μg to the user.    -   17—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 400 μg to the user.    -   18—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is formulated        to provide a dose of about 500 μg to the user.    -   19—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 10 μg/ml to 2 mg/ml.    -   20—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 50 μg/ml to 2 mg/ml.    -   21—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 100 μg/ml to 1 mg/ml.    -   22—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 10 μg/ml to 1 mg/ml.    -   23—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 50 μg/ml to 750 μg/ml.    -   24—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 100 μg/ml to 500 μg/ml.    -   25—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of between 200 μg/ml to 500 μg/ml.    -   26—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of 300 μg/ml±20%.    -   27—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof is at a        concentration of 300 μg/ml±10%.    -   28—The pharmaceutical formulation of any one of the preceding        embodiments comprising a surfactant.    -   29—The pharmaceutical formulation of any one of the preceding        embodiments comprising a non-ionic surfactant.    -   30—The pharmaceutical formulation of any one of the preceding        embodiments comprising polysorbate 20.    -   31—The pharmaceutical formulation of any one of the preceding        embodiments comprising polysorbate 40.    -   32—The pharmaceutical formulation of any one of the preceding        comprising polysorbate 60.    -   33—The pharmaceutical formulation of any one of the preceding        embodiments comprising polysorbate 65.    -   34—The pharmaceutical formulation of any one of the preceding        embodiments comprising polysorbate 80.    -   35—The pharmaceutical formulation of any one of the preceding        embodiments comprising Triton X-100.    -   36—The pharmaceutical formulation of any one of the preceding        embodiments comprising poloxamer.    -   37—The pharmaceutical formulation of any one of the preceding        embodiments comprising pluronic F-68.    -   38—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the buffering agent is acetate, citrate,        histidine, phosphate, succinate, maleate or tromethamine.    -   39—The pharmaceutical formulation of any one of the preceding        embodiments comprising citrate as buffering agent.    -   40—The pharmaceutical formulation of any one of the preceding        embodiments comprising histidine-HCl as buffering agent.    -   41—The pharmaceutical formulation of any one of the preceding        embodiments comprising sodium phosphate as buffering agent.    -   42—The pharmaceutical formulation of any one of the preceding        embodiments comprising potassium phosphate as buffering agent.    -   43—The pharmaceutical formulation of any one of the preceding        embodiments comprising maleate as buffering agent.    -   44—The pharmaceutical formulation of any one of the preceding        embodiments comprising tromethamine as buffering agent.    -   45—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the tonicity agent is selected from the        group consisting of dextrose, glucose, glycerin, glycerol,        mannitol, sodium chloride, sodium sulfate, sorbitol, sucrose,        trehalose and potassium chloride.    -   46—The pharmaceutical formulation of any one of the preceding        embodiments comprising glycerol as tonicity agent.    -   47—The pharmaceutical formulation of any one of the preceding        embodiments comprising mannitol as tonicity agent.    -   48—The pharmaceutical formulation of any one of the preceding        embodiments comprising sucrose as tonicity agent.    -   49—pharmaceutical formulation of any one of the preceding        embodiments comprising trehalose as tonicity agent.    -   50—The pharmaceutical formulation of any one of the preceding        embodiments comprising amino acids excipients.    -   51—The pharmaceutical formulation of any one of the preceding        embodiments comprising alanine as excipient.    -   52—The pharmaceutical formulation of any one of the preceding        embodiments comprising arginine as excipient.    -   53—The pharmaceutical formulation of any one of the preceding        embodiments comprising glycine as excipient.    -   54—The pharmaceutical formulation of any one of the preceding        embodiments comprising leucine as excipient.    -   55—The pharmaceutical formulation of any one of the preceding        embodiments comprising methionine as excipient.    -   56—The pharmaceutical formulation of any one of the preceding        embodiments comprising proline as excipient.    -   57—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is at a pH        of between 6.5±0.5 to 7.5±0.5.    -   58—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is at a pH        of between 6.0±0.15 to 7.5±0.15.    -   59—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is at a pH        of between 6.5±0.5 to 7.0±0.5.    -   60—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is at a pH        of between 6.5±0.15 to 7.0±0.15.    -   61—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the formulation is at pH 6.5±0.5.    -   62—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the formulation is at pH 6.5±0.15.    -   63—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the formulation is at pH 7.0±0.5.    -   64—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the formulation is at pH 7.0±0.15.    -   65—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the formulation is at pH 7.5±0.5.    -   66—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the buffer is at concentration of from        about 1 mM to about 50 mM.    -   67—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the tonicity agent is at concentration of        from about 10 mM to about 500 mM.    -   68—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the tonicity agent is at concentration of        from about 1% to about 20% (w/v).    -   69—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the surfactant is at a concentration of        from about 0.001 to about 0.1% (w/v),    -   70—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the amino acid excipient at a concentration        of from about 0.5 mM to about 300 mM.    -   71—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a) a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 10 μg/ml        to about 1 mg/ml, b) a citrate buffer at a concentration of from        about 1 mM to about 50 mM, c) glycerol at a concentration of        from about 10 mM to about 500 mM, mannitol at a concentration of        from about 10 mM to about 500 mM, sucrose at a concentration of        from about 1% to about 20%, trehalose at a concentration of from        about 1% to about 20% or NaCl at a concentration of from about        50 mM to about 150 mM, d) optionally polysorbate at a        concentration of from about 0.001 to about 0.1% (w/v), and e)        optionally an amino acid excipient at a concentration of about        0.5 mM to about 300 mM.    -   72—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a) a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 10 μg/ml        to about 1 mg/ml, b) a histidine buffer at a concentration of        from about 1 mM to about 50 mM, c) glycerol at a concentration        of from about 10 mM to about 500 mM, mannitol at a concentration        of from about 10 mM to about 500 mM, sucrose at a concentration        of from about 1% to about 20%, trehalose at a concentration of        from about 1% to about 20% or NaCl at a concentration of from        about 50 mM to about 150 mM d) optionally polysorbate at a        concentration of from about 0.001 to about 0.1% (w/v), and e)        optionally an amino acid excipient at a concentration of about        0.5 mM to about 300 mM.    -   73—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a) a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 10 μg/ml        to about 1 mg/ml, b) a maleate buffer at a concentration of from        about 1 mM to about 50 mM, c) glycerol at a concentration of        from about 10 mM to about 500 mM, mannitol at a concentration of        from about 10 mM to about 500 mM, sucrose at a concentration of        from about 1% to about 20%, trehalose at a concentration of from        about 1% to about 20% or NaCl at a concentration of from about        50 mM to about 150 mM, d) optionally polysorbate at a        concentration of from about 0.001 to about 0.1% (w/v), and e)        optionally an amino acid excipient at a concentration of about        0.5 mM to about 300 mM.    -   74—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a) a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 10 μg/ml        to about 1 mg/ml, b) a phosphate buffer at a concentration of        from about 1 mM to about 50 mM, c) glycerol at a concentration        of from about 10 mM to about 500 mM, mannitol at a concentration        of from about 10 mM to about 500 mM, sucrose at a concentration        of from about 1% to about 20%, trehalose at a concentration of        from about 1% to about 20% or NaCl at a concentration of from        about 50 mM to about 150 mM d) optionally polysorbate at a        concentration of from about 0.001 to about 0.1% (w/v), and e)        optionally an amino acid excipient at a concentration of about        0.5 mM to about 300 mM.    -   75—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a GLP-1 or a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, citrate at a concentration of from about 7.5        mM to about 12.5 mM±10% and glycerol at a concentration of from        about 150 mM to about 300 mM±10%, mannitol at a concentration of        from about 150 mM to about 300 mM±10%, sucrose at a        concentration of from about 5% to about 15%±10% or trehalose at        a concentration of from about 5% to about 15%±10%.    -   76—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a GLP-1 or a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of from        about 7.5 mM to about 12.5 mM±10% and glycerol at a        concentration of from about 150 mM to about 300 mM±10%, mannitol        at a concentration of from about 150 mM to about 300 mM±10%,        sucrose at a concentration of from about 5% to about 15%±10% or        trehalose at a concentration of from about 5% to about 15%±10%.    -   77—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a GLP-1 or a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of from        about 7.5 mM to about 12.5 mM±10% and glycerol at a        concentration of from about 150 mM to about 300 mM±10%, mannitol        at a concentration of from about 150 mM to about 300 mM±10%        sucrose at a concentration of from about 5% to about 15%±10%, or        trehalose at a concentration of from about 5% to about 15%±10%.    -   78—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a GLP-1 or a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, potassium phosphate at a concentration of        from about 7.5 mM to about 12.5 mM±10% and glycerol at a        concentration of from about 150 mM to about 300 mM±10%, mannitol        at a concentration of from about 150 mM to about 300 mM±10%,        sucrose at a concentration of from about 5% to about 15%±10% or        trehalose at a concentration of from about 5% to about 15%±10%.    -   79—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 or 6.0±0.15 to 7.5±0.15 and including a GLP-1 or a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, maleate at a concentration of from about 7.5        mM to about 12.5 mM±10% and glycerol at a concentration of from        about 150 mM to about 300 mM±10%, mannitol at a concentration of        from about 150 mM to about 300 mM±10%, sucrose at a        concentration of from about 5% to about 15%±10% or trehalose at        a concentration of from about 5% to about 15%±10%.    -   80—A pharmaceutical formulation at a pH of 6.5±0.5 to 7.5±0.5        (e.g., 6.5±0.15 to 7.0±0.15 etc.) and including a GLP-1 analogue        comprising or consisting of the amino acid sequence set forth in        SEQ ID NO:3 at a concentration of from about 100 μg/ml to about        500 μg/ml, sodium citrate at a concentration of about 10 mM±10%,        a tonicity agent such as glycerol at a concentration of from        about 165 mM to about 275 mM±10%, sucrose at a concentration of        from about 5% to about 15%, trehalose at a concentration of from        about 5% to about 15% or NaCl at a concentration of from about        50 mM to about 150 mM, a surfactant such as polysorbate at a        concentration of about 0.005 to 0.05% (w/v)±10% and an amino        acid excipient such as methionine at a concentration of about 5        mM to about 25 mM±10%, proline at a concentration of from about        25 mM to about 75 mM±10% or arginine ata concentration of from        about 50 mM to about 150 mM±10%.    -   81—A pharmaceutical formulation at a pH of 6.5±0.5 to 7.5±0.5        (e.g., 6.5±0.15 to 7.0±0.15 etc.) and including a GLP-1 analogue        comprising or consisting of the amino acid sequence set forth in        SEQ ID NO:3 at a concentration of from about 100 μg/ml to about        500 μg/ml, histidine HCl at a concentration of about 10 mM±10%,        a tonicity agent such as glycerol at a concentration of from        about 165 mM to about 275 mM±10%, sucrose at a concentration of        from about 5% to about 15%, trehalose at a concentration of from        about 5% to about 15% or NaCl at a concentration of from about        50 mM to about 150 mM, a surfactant such as polysorbate at a        concentration of about 0.005 to 0.05% (w/v)±10% and an amino        acid excipient such as methionine at a concentration of about 5        mM to about 25 mM±10%, proline at a concentration of from about        25 mM to about 75 mM±10% or arginine at a concentration of from        about 50 mM to about 150 mM±10%.    -   82—A pharmaceutical formulation at a pH of 6.5±0.5 to 7.5±0.5        (e.g., 6.5±0.15 to 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, maelate at a concentration of about 10        mM±10%, a tonicity agent such as glycerol at a concentration of        from about 165 mM to about 275 mM±10%, sucrose at a        concentration of from about 5% to about 15%, trehalose at a        concentration of from about 5% to about 15% or NaCl at a        concentration of from about 50 mM to about 150 mM, a surfactant        such as polysorbate at a concentration of about 0.005 to 0.05%        (wv)±10% and an amino acid excipient such as methionine at a        concentration of about 5 mM to about 25 mM±10%, proline at a        concentration of from about 25 mM to about 75 mM±10% or arginine        at a concentration of from about 50 mM to about 150 mM±10%.    -   83—A pharmaceutical formulation at a pH of 6.5±0.5 to 7.5±0.5        (e.g., 6.5±0.15 to 7.0±0.15 etc.) and including a GLP-1 analogue        comprising or consisting of the amino acid sequence set forth in        SEQ ID NO:3 at a concentration of from about 100 μg/ml to about        500 μg/ml, sodium phosphate at a concentration of about 10        mM±10%, a tonicity agent such as glycerol at a concentration of        from about 165 mM to about 275 mM±10%, sucrose at a        concentration of from about 5% to about 15%, trehalose at a        concentration of from about 5% to about 15% or NaCl at a        concentration of from about 50 mM to about 150 mM, a surfactant        such as polysorbate at a concentration of about 0.005 to 0.05%        (w/v)±10% and an amino acid excipient such as methionine at a        concentration of about 5 mM to about 25 mM±10%, proline at a        concentration of from about 25 mM to about 75 mM±10% or arginine        at a concentration of from about 50 mM to about 150 mM±10%.    -   84—A pharmaceutical formulation at a pH of 6.5±0.5 to 7.5±0.5        (e.g., 6.5±0.15 to 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, potassium phosphate at a concentration of        about 10 mM±10%, a tonicity agent such as glycerol at a        concentration of from about 165 mM to about 275 mM±10%, sucrose        at a concentration of from about 5% to about 15%, trehalose at a        concentration of from about 5% to about 15% or NaCl at a        concentration of from about 50 mM to about 150 mM, a surfactant        such as polysorbate at a concentration of about 0.005 to 0.05%        (w/v)±10% and an amino acid excipient such as methionine at a        concentration of about 5 mM to about 25 mM±10%, proline at a        concentration of from about 25 mM to about 75 mM±10% or arginine        at a concentration of from about 50 mM to about 150 mM±10%.    -   85—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, and mannitol at a concentration of about 165 mM to about        275 mM±10%, sucrose at a concentration of about 5% to about        15%±10% or trehalose at a concentration of about 5% to about        15%±10%.    -   86—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 165 mM to about 275        mM±10%, and polysorbate-20 at a concentration of about 0.01 to        0.03% (w/v)±10%.    -   87—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 165 mM±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   88—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 275 mM±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   89—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., (e.g., 7.0±0.5, 7.0±0.15 etc.) and including a        GLP-1 analogue comprising or consisting of the amino acid        sequence set forth in SEQ ID NO:3 at a concentration of from        about 100 μg/ml to about 500 μg/ml, histidine-HCl at a        concentration of about 10 mM±10%, mannitol at a concentration of        about 275 mM±10%, polysorbate-20 at a concentration of about        0.01 to 0.03% (w/v)±10% and methionine at a concentration of        about 5 mM to about 25 mM±10%.    -   90—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 165 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   91—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 275 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   92—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, mannitol at a concentration of about 165 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   93—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 5% to about 15%±10%,        and polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   94—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 8%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   95—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 10%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   96—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   97—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   98—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   99—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, sucrose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   100—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 5% to about        15%±10%, and polysorbate-20 at a concentration of about 0.01 to        0.03% (w/v)±10%.    -   101—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 8%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   102—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 10%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   103—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   104—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   105—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM 10%.    -   106—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 7.0±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, histidine-HCl at a concentration of about 10        mM±10%, trehalose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   107—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, and mannitol at a concentration of about 165 mM to        about 275 mM±10% sucrose at a concentration of about 5% to about        15%±10% or trehalose at a concentration of about 5% to about        15%±10%.    -   108—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.(=) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 165 mM to about        275 mM±10%, and polysorbate-20 at a concentration of about 0.01        to 0.03% (w/v)±10%.    -   109—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 165 mM±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   110—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about about 275        mM±10%, and polysorbate-20 at a concentration of about 0.01 to        0.03% (w/v)±10%.    -   111—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 275 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   112—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 165 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   113—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 275 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   114—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, mannitol at a concentration of about 165 mM±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   115—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 5% to about        15%±10%, and polysorbate-20 at a concentration of about 0.01 to        0.03% (w/v)±10%.    -   116—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 8%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   117—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 10%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   118—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   119—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   120—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   121—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, sucrose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   122—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 5% to about        15%±10%, and polysorbate-20 at a concentration of about 0.01 to        0.03% (w/v)±10%.    -   123—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 8%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   124—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 10%±10%, and        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10%.    -   125—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   126—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and methionine at a concentration of about 5 mM to        about 25 mM±10%.    -   127—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 8%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   128—A pharmaceutical formulation at a pH of about 6.5±0.5 to        7.5±0.5 (e.g., 7.0±0.5, 6.5±0.15, etc.) and including a GLP-1        analogue comprising or consisting of the amino acid sequence set        forth in SEQ ID NO:3 at a concentration of from about 100 μg/ml        to about 500 μg/ml, sodium phosphate at a concentration of about        10 mM±10%, trehalose at a concentration of about 10%±10%,        polysorbate-20 at a concentration of about 0.01 to 0.03%        (w/v)±10% and proline at a concentration of about 25 mM to about        75 mM±10%.    -   129—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is isotonic with blood        and/or serum.    -   130—The pharmaceutical formulation of any one of the preceding        embodiments wherein the peptide concentration is ±20% the        initial concentration (at preparation) after one or more stress        conditions.    -   131—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation has a low concentration of        subvisible particles.    -   132—The pharmaceutical formulation of any one of the preceding        embodiments wherein the concentration of subvisible particles        (cumulative particle counts) as measured by micro-flow imaging        is 1000 or below for particles ≥2 μm, 275 or below for particles        ≥5 μm, 100 or below for particles ≥10 μm and/or 30 or below for        particles 25 μm at T−1m_25° C. and/or T−1m_40° C.    -   133—The pharmaceutical formulation of any one of the preceding        embodiments wherein the concentration of subvisible particles        (cumulative particle counts) as measured by micro-flow imaging        is 500 or below for particles ≥2 μm, 200 or below for particles        ≥5 μm, 50 or below for particles 0 μm and/or 10 or below for        particles 25 μm at T−3m_5° C. and/or T−3m_25° C.    -   134—The pharmaceutical formulation of any one of the preceding        embodiments wherein the monomer content as measured by HP-SEC is        95% or higher at T−1m_5° C., 95% or higher at T−1m_25° C., 92%        or higher at T−1m_40° C., 95% or higher at T−3m_5° C., 95% or        higher at T−3m_25° C., and/or 92% or higher at T−3m_40° C.    -   135—The pharmaceutical formulation of any one of the preceding        embodiments wherein the monomer content as measured by HP-SEC is        at least 92%, at least 93%, at least 94% or at least 95% or        higher under one or more stress conditions.    -   136—The pharmaceutical formulation of any one of the preceding        embodiments wherein the aggregate content as measured by HP-SEC        is 5% or less, 4% or less, 3% or less. 2% or less, 1% or less at        T−1m_5° C., T−1m_25° C., T−3m_5° C. and/or T−3m_25° C.    -   137—The pharmaceutical formulation of any one of the preceding        embodiments wherein the aggregate content as measured by HP-SEC        is 5% or less, 4% or less, 3% or less. 2% or less, 1% or less        under one or more stress conditions.    -   138—The pharmaceutical formulation of any one of the preceding        embodiments provided in a single-dose or multi-dose container.    -   139—The pharmaceutical formulation of any one of the preceding        embodiments, in a unit-dose vial, multi-dose vial, cartridge or        pre-filled syringe.    -   140—The pharmaceutical formulation of any one of the preceding        embodiments in a unit-dose vial, multi-dose vial, cartridge or        pre-filled syringe that includes, for example, from about 20 μg        to about 1 mg of the GLP-1 or GLP-1 analogue or ROSE-010.    -   141—The pharmaceutical formulation of any one of the preceding        embodiments, in a unit-dose vial, multi-dose vial, cartridge or        pre-filled syringe that includes for example, a dose of at least        150 μg of the GLP-1 or GLP-1 analogue or ROSE-010.    -   142—The pharmaceutical formulation of any one of the preceding        embodiments wherein sodium acetate is used at a concentration of        10 mM.    -   143—The pharmaceutical formulation of any one of the preceding        embodiments wherein sodium citrate is used at a concentration of        10 mM.    -   144—The pharmaceutical formulation of any one of the preceding        embodiments wherein Histidine-HCl is used at a concentration of        10 mM.    -   145—The pharmaceutical formulation of any one of the preceding        embodiments wherein sodium maleate is used at a concentration of        10 mM.    -   146—The pharmaceutical formulation of any one of the preceding        embodiments wherein sodium phosphate is used at a concentration        of 10 mM.    -   147—The pharmaceutical formulation of any one of the preceding        embodiments wherein glycerol is used at a concentration of 165        mM.    -   148—The pharmaceutical formulation of any one of the preceding        embodiments wherein glycerol is used at a concentration of 275        mM.    -   149—The pharmaceutical formulation of any one of the preceding        embodiments wherein mannitol is used at a concentration of 3%.    -   150—The pharmaceutical formulation of any one of the preceding        embodiments wherein mannitol is used at a concentration of 5%.    -   151—The pharmaceutical formulation of any one of the preceding        embodiments wherein NaCl is used at a concentration of 50 mM.    -   152—The pharmaceutical formulation of any one of the preceding        embodiments wherein NaCl is used at a concentration of 130 mM.    -   153—The pharmaceutical formulation of any one of the preceding        embodiments wherein NaCl is used at a concentration of 150 mM.    -   154—The pharmaceutical formulation of any one of the preceding        embodiments wherein sucrose is used at a concentration of 5%.    -   155—The pharmaceutical formulation of any one of the preceding        embodiments wherein sucrose is used at a concentration of 8%.    -   156—The pharmaceutical formulation of any one of the preceding        embodiments wherein sucrose is used at a concentration of 10%.    -   157—The pharmaceutical formulation of any one of the preceding        embodiments wherein trehalose is used at a concentration of 5%.    -   158—The pharmaceutical formulation of any one of the preceding        embodiments wherein trehalose is used at a concentration of 8%.    -   159—The pharmaceutical formulation of any one of the preceding        embodiments wherein trehalose is used at a concentration of 10%.    -   160—The pharmaceutical formulation of any one of the preceding        embodiments wherein arginine-HCl is used at a concentration of        50 mM.    -   161—The pharmaceutical formulation of any one of the preceding        embodiments wherein arginine-HCl is used at a concentration of        100 mM.    -   162—The pharmaceutical formulation of any one of the preceding        embodiments wherein methionine is used at a concentration of 5        mM.    -   163—The pharmaceutical formulation of any one of the preceding        embodiments wherein methionine is used at a concentration of 10        mM.    -   164—The pharmaceutical formulation of any one of the preceding        embodiments wherein methionine is used at a concentration of 15        mM.    -   165—The pharmaceutical formulation of any one of the preceding        embodiments wherein proline is used at a concentration of 25 mM.    -   166—The pharmaceutical formulation of any one of the preceding        embodiments wherein proline is used at a concentration of 50 mM.    -   167—The pharmaceutical formulation of any one of the preceding        embodiments wherein proline is used at a concentration of 75 mM.    -   168—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-20 at a        concentration of 0.005% (w/v).    -   169—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-20 at a        concentration of 0.02% (w/v).    -   170—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-20 at a        concentration of 0.05% (w/v).    -   171—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-60 at a        concentration of 0.005% (w/v).    -   172—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-60 at a        concentration of 0.02% (w/v).    -   173—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-60 at a        concentration of 0.05% (w/v).    -   174—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-80 at a        concentration of 0.005% (w/v).    -   175—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-80 at a        concentration of 0.02% (w/v).    -   176—The pharmaceutical formulation of any one of the preceding        embodiments wherein the polysorbate is polysorbate-80 at a        concentration of 0.05% (w/v).    -   177—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 6.5.    -   178—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 6.6.    -   179—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 6.7.    -   180—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 6.8.    -   181—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 6.9.    -   182—The pharmaceutical formulation of any one of the preceding        embodiments wherein the formulation is at pH 7.0.    -   183—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof comprises or        consists of the amino acid sequence set forth in SEQ ID NO:3 at        a concentration of 300 μg/ml±20%.    -   184—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the GLP-1 or analogue thereof comprises or        consists of the amino acid sequence set forth in SEQ ID NO:3 at        a concentration of 300 μg/ml±10%.    -   185—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is liquid.    -   186—The pharmaceutical formulation of any one of the preceding        embodiments, wherein the pharmaceutical formulation is stable.    -   187—A pre-filled syringe comprising the pharmaceutical        formulation of any one of the preceding embodiments.    -   188—A pre-filled syringe comprising the pharmaceutical        formulation of any one of the preceding embodiments that        comprises from about 20 μg to about 1 mg of the GLP-1 or GLP-1        analogue or ROSE-010.    -   189—A pre-filled syringe comprising the pharmaceutical        formulation of any one of the preceding embodiments that        comprises a dose of at least 150 μg of the GLP-1 or GLP-1        analogue or ROSE-010.    -   190—A pre-filled syringe comprising the pharmaceutical        formulation of any one of the preceding embodiments, wherein the        GLP-1 or GLP-1 analogue or ROSE-010 thereof is at a        concentration of 300 μg/ml±10%.    -   191—A method of treating a disorder or condition in which        administration of a GLP-1 or GLP-1 analogue or ROSE-010 is        indicated, that comprises administering the pharmaceutical        formulation of any one of the preceding embodiments to an        individual in need.    -   192—A method of treating irritable bowel syndrome (IBS) that        comprises administering the pharmaceutical formulation of any        one of the preceding embodiments to an individual in need.    -   193—A method of treating constipation predominant IBS (IBS-C)        that comprises administering the pharmaceutical formulation of        any one of the preceding embodiments to an individual in need.    -   194—A method of treating diarrhea predominant IBS (IBS-D) that        comprises administering the pharmaceutical formulation of any        one of the preceding embodiments to an individual in need.    -   195—A method of treating mixed or alternating type IBS (IBS-M)        that comprises administering the pharmaceutical formulation of        any one of the preceding embodiments to an individual in need.    -   196—A method of treating diabetes, ischemia, reperfused tissue        injury, dyslipidemia, diabetic cardiomyopathy, myocardial        infarction, acute coronary syndrome, obesity, catabolic changes        after surgery, hyperglycemia, irritable bowel syndrome, stroke,        neurodegenerative disorders, memory and learning disorders,        islet cell transplant functional dyspepsia and/or regenerative        therapy that comprises administering the pharmaceutical        formulation of any one of the preceding embodiments to an        individual in need.    -   197—The method of any one of the preceding embodiments, wherein        the pharmaceutical formulation is administered subcutaneously.    -   198—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 20 μg to about 1 mg.    -   199—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 30 μg to about 300 μg daily.    -   200—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 50 μg.    -   201—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 100 μg.    -   202—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 150 μg.    -   203—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 200 μg.    -   204—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 300 μg.    -   205—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 400 μg.    -   206—The method of any one of the preceding embodiments, wherein        the GLP-1 or GLP-1 analogue or ROSE-010 is administered at a        dose of about 500 μg.    -   207—The method of any one of the preceding embodiments, wherein        the pharmaceutical formulation is administered during acute pain        associated with IBS.    -   208—The method of any one of the preceding embodiments, wherein        the pharmaceutical formulation is administered during acute pain        associated with IBS-C, IBS-D or IBS-M.    -   209—The pharmaceutical formulation of any one of the preceding        embodiments for use in a method of treatment of any one of the        preceding embodiments.

The following examples are provided to further illustrate aspects andembodiments of the disclosure. These examples are non-limiting andshould not be construed as limiting any aspects and embodiments of thedisclosure.

EXAMPLES Example 1: Peptide Material, Formulations and Methods

The solid ROSE-010 acetate salt (net peptide content of 88.12%; purity(HPLC) of 96.1%) is aliquoted into 200 mg aliquots and stored at −20° C.Aliquots used in experimentations re thawed at room temperature.

Stock solutions of each component (buffer, tonicity agent, excipientand/or surfactant etc.) are prepared, filtered and are either usedimmediately or stored at room temperature or between 2-8° C.

Stock formulations at 1 mg/ml are prepared by using concentrated peptidesolution spiked in water or into stock solutions of buffer, excipientsand/or tonicity agent and filled up with water. If needed, solubility ofthe ROSE-010 (acetate salt) may be increased by the addition of an acidsuch as HCl. If needed, the pH of the samples is adjusted.

The peptide concentration is confirmed by UV spectroscopy (λ=280 nm).The final peptide concentration in the formulations tested is set at 300μg/ml (Table 4). Corresponding placebo formulations (without peptide)are made by using the same buffer, excipients and/or tonicity agents.

The samples are filtered under laminar air-flow (LAF), by using a0.22-μm syringe PVDF filter unit. Subsequently, 1.5 ml of the filteredformulations are filled into washed and sterilized 2R glass vials.

The following reagents (USP or Ph. Eur. grade) are used:

Name   L-Histidine Hydrochloride Monohydrate L-Histidine D-MannitolL-Methionine Tween ® 20 (Polysorbate) NF, Multi-compendial Sodiumacetate, Trihydrate Tri-Sodium citrate, Dihydrate Sodium dihydrogenphosphate, Dihydrate Di-sodium hydrogen phosphate, Dihydrate NaCl,D-Sucrose D-Trehalose, Dihydrate L-Arginine L-Proline Glycerol

Several formulations are tested to determine the effect of pH, bufferingagents, tonicity agents, excipients and surfactant on the stability ofROSE-010 peptide. Representative formulations are illustrated in Tables1A-1E and Table 4. It is to be understood herein that in Tables 1A-1Dand Table 4, the pH is provided for illustrative purposes only and mayvary without affecting the stability of the formulation. For example,formulations containing histidine-HCl or sodium phosphate may have a pHranging from 6.5±0.5 to 7.5±0.5, including for example, a pH rangingfrom 6.5±0.5 to 7.0±0.5, a pH of 6.5±0.5, a pH of 7.0±0.5, a pH of7.5±0.5 etc.

TABLE 1A List of histidine buffer-based formulations pH Buffer Tonicityagent Excipient Surfactant 6.5 ± 0.15 10 mM histidine-HCl  5% sucrose — 0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl  5% sucrose 10 mM 0.02% (w/v) PS20 methionine 6.5 ± 0.15 10 mM histidine-HCl  5% sucrose50 mM proline  0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl  8%sucrose —  0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl  8% sucrose10 mM  0.02% (w/v) PS20 methionine 6.5 ± 0.15 10 mM histidine-HCl  8%sucrose 50 mM proline  0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl10% sucrose —  0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl 10%sucrose 10 mM  0.02% (w/v) PS20 methionine 6.5 ± 0.15 10 mMhistidine-HCl 10% sucrose 50 mM proline  0.02% (w/v) PS20 6.5 ± 0.15 10mM histidine-HCl 165 mM mannitol —  0.02% (w/v) PS20 6.5 ± 0.15 10 mMhistidine-HCl 165 mM mannitol 10 mM  0.02% (w/v) PS20 methionine 6.5 ±0.15 10 mM histidine-HCl 165 mM mannitol 50 mM proline  0.02% (w/v) PS206.5 ± 0.15 10 mM histidine-HCl 275 mM mannitol —  0.02% (w/v) PS20 6.5 ±0.15 10 mM histidine-HCl 275 mM mannitol 10 mM  0.02% (w/v) PS20methionine 6.5 ± 0.15 10 mM histidine-HCl 275 mM mannitol 50 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  5% sucrose —  0.02%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  5% sucrose 10 mM  0.02% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  5% sucrose 50 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose —  0.02%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose  5 mM 0.005% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 10 mM 0.005%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 15 mM0.005% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose5 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  8%sucrose 10 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl  8% sucrose 15 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.1510 mM histidine-HCl  8% sucrose  5 mM  0.05% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl  8% sucrose 10 mM  0.05% (w/v) PS20 methionine7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 15 mM  0.05% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 15 mM  0.02% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 25 mM proline0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 50 mMproline 0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose 75mM proline 0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8% sucrose25 mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl  8%sucrose 50 mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8% sucrose 75 mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl  8% sucrose 25 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10mM histidine-HCl  8% sucrose 50 mM proline  0.05% (w/v) PS20 7.0 ± 0.1510 mM histidine-HCl  8% sucrose 75 mM proline  0.05% (w/v) PS20 7.0 ±0.15 10 mM histidine-HCl 10% sucrose —  0.02% (w/v) PS20 7.0 ± 0.15 10mM histidine-HCl 10% sucrose  5 mM 0.005% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl 10% sucrose 10 mM 0.005% (w/v) PS20 methionine7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 15 mM 0.005% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose  5 mM  0.02% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 10 mM  0.02%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 15 mM 0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 5 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10%sucrose 10 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 10% sucrose 15 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.1510 mM histidine-HCl 10% sucrose 25 mM proline 0.005% (w/v) PS20 7.0 ±0.15 10 mM histidine-HCl 10% sucrose 50 mM proline 0.005% (w/v) PS20 7.0± 0.15 10 mM histidine-HCl 10% sucrose 75 mM proline 0.005% (w/v) PS207.0 ± 0.15 10 mM histidine-HCl 10% sucrose 25 mM proline  0.02% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 50 mM proline  0.02%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 75 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 25 mMproline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 50mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose75 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mMmannitol — 0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mMmannitol (3% —  0.02% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl165 mM mannitol (3% —  0.05% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mMhistidine-HCl 165 mM mannitol (3%  5 mM 0.005% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 10 mM0.005% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 165mM mannitol (3% 15 mM 0.005% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10mM histidine-HCl 165 mM mannitol (3%  5 mM  0.02% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 10 mM 0.02% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 165mM mannitol (3% 15 mM  0.02% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10mM histidine-HCl 165 mM mannitol (3%  5 mM  0.05% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 10 mM 0.05% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 165mM mannitol (3% 15 mM  0.05% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10mM histidine-HCl 165 mM mannitol (3% 25 mM proline 0.005% (w/v) PS20(w/v)) 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 50 mM proline0.005% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol(3% 75 mM proline 0.005% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mMhistidine-HCl 165 mM mannitol (3% 25 mM proline  0.02% (w/v) PS20 (w/v))7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 50 mM proline  0.02%(w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 75mM proline  0.02% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 165mM mannitol (3% 25 mM proline  0.05% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mMhistidine-HCl 165 mM mannitol (3% 50 mM proline  0.05% (w/v) PS20 (w/v))7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol (3% 75 mM proline  0.05%(w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% — —(w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% — 0.005% (w/v)PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% —  0.02%(w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% — 0.05% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol(5%  5 mM 0.005% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mMhistidine-HCl 275 mM mannitol (5% 10 mM 0.005% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 15 mM0.005% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 275mM mannitol (5%  5 mM  0.02% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10mM histidine-HCl 275 mM mannitol (5% 10 mM  0.02% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 15 mM 0.02% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 275mM mannitol (5% 5 mM  0.05% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10mM histidine-HCl 275 mM mannitol (5% 10 mM  0.05% (w/v) PS20 (w/v))methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 15 mM 0.05% (w/v) PS20 (w/v)) methionine 7.0 ± 0.15 10 mM histidine-HCl 275mM mannitol (5% 25 mM proline 0.005% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mMhistidine-HCl 275 mM mannitol (5% 50 mM proline 0.005% (w/v) PS20 (w/v))7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 75 mM proline 0.005%(w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 25mM proline  0.02% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275mM mannitol (5% 50 mM proline  0.02% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mMhistidine-HCl 275 mM mannitol (5% 75 mM proline  0.02% (w/v) PS20 (w/v))7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 25 mM proline  0.05%(w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275 mM mannitol (5% 50mM proline  0.05% (w/v) PS20 (w/v)) 7.0 ± 0.15 10 mM histidine-HCl 275mM mannitol (5% 75 mM proline  0.05% (w/v) PS20 (w/v))

TABLE 1B List of histidine buffer-based formulations pH Buffer Tonicityagent Excipient Surfactant 6.5 ± 0.15 10 mM histidine-HCl 5% trehalose — 0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl 5% trehalose 10 mM 0.02% (w/v) PS20 methionine 6.5 ± 0.15 10 mM histidine-HCl 5% trehalose50 mM  0.02% (w/v) PS20 proline 6.5 ± 0.15 10 mM histidine-HCl 5%trehalose 50 mM  0.02% (w/v) PS20 arginine-HCl 6.5 ± 0.15 10 mMhistidine-HCl 8% trehalose —  0.02% (w/v) PS20 6.5 ± 0.15 10 mMhistidine-HCl 8% trehalose 10 mM  0.02% (w/v) PS20 methionine 6.5 ± 0.1510 mM histidine-HCl 8% trehalose 50 mM proline  0.02% (w/v) PS20 6.5 ±0.15 10 mM histidine-HCl 8% trehalose 50 mM  0.02% (w/v) PS20arginine-HCl 6.5 ± 0.15 10 mM histidine-HCl 10% trehalose  — — 6.5 ±0.15 10 mM histidine-HCl 10% trehalose  —  0.02% (w/v) PS20 6.5 ± 0.1510 mM histidine-HCl 10% trehalose  10 mM  0.02% (w/v) PS20 methionine6.5 ± 0.15 10 mM histidine-HCl 10% trehalose  50 mM proline  0.02% (w/v)PS20 6.5 ± 0.15 10 mM histidine-HCl 10% trehalose  50 mM  0.02% (w/v)PS20 arginine-HCl 6.5 ± 0.15 10 mM histidine-HCl 165 mM glycerol — 0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl 165 mM glycerol 10 mM 0.02% (w/v) PS20 methionine 6.5 ± 0.15 10 mM histidine-HCl 165 mMglycerol 50 mM proline  0.02% (w/v) PS20 6.5 ± 0.15 10 mM histidine-HCl165 mM glycerol 50 mM  0.02% (w/v) PS20 arginine-HCl 6.5 ± 0.15 10 mMhistidine-HCl 275 mM glycerol —  0.02% (w/v) PS20 6.5 ± 0.15 10 mMhistidine-HCl 275 mM glycerol 10 mM  0.02% (w/v) PS20 methionine 6.5 ±0.15 10 mM histidine-HCl 275 mM glycerol 50 mM proline  0.02% (w/v) PS206.5 ± 0.15 10 mM histidine-HCl 275 mM glycerol 50 mM  0.02% (w/v) PS20arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl 5% trehalose —  0.02% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 5% trehalose 10 mM  0.02% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 5% trehalose 50 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 5% trehalose 50 mM 0.02% (w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl 8%trehalose —  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8%trehalose  5 mM 0.005% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 8% trehalose 10 mM 0.005% (w/v) PS20 methionine 7.0 ± 0.1510 mM histidine-HCl 8% trehalose 15 mM 0.005% (w/v) PS20 methionine 7.0± 0.15 10 mM histidine-HCl 8% trehalose  5 mM  0.02% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 10 mM  0.02%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 15 mM 0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 5 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 8%trehalose 10 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 8% trehalose 15 mM  0.05% (w/v) PS20 methionine 7.0 ± 0.1510 mM histidine-HCl 8% trehalose 15 mM  0.02% (w/v) PS20 methionine 7.0± 0.15 10 mM histidine-HCl 8% trehalose 25 mM proline 0.005% (w/v) PS207.0 ± 0.15 10 mM histidine-HCl 8% trehalose 50 mM proline 0.005% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 75 mM proline 0.005%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 25 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 50 mMproline  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose 75mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8% trehalose25 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 8%trehalose 50 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl8% trehalose 75 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 8% trehalose 50 mM  0.02% (w/v) PS20 arginine-HCl 7.0 ±0.15 10 mM histidine-HCl — 150 mM   0.02% (w/v) PS20 arginine-HCl 7.0 ±0.15 10 mM histidine-HCl 10% trehalose  — — 7.0 ± 0.15 10 mMhistidine-HCl 10% trehalose  —  0.02% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 10% trehalose   5 mM 0.005% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl 10% trehalose  10 mM 0.005% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  15 mM 0.005%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose   5mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10%trehalose  10 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 10% trehalose  15 mM  0.02% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl 10% trehalose   5 mM  0.05% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  10 mM  0.05%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  15mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 10%trehalose  25 mM proline 0.005% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 10% trehalose  50 mM proline 0.005% (w/v) PS20 7.0 ± 0.1510 mM histidine-HCl 10% trehalose  75 mM proline 0.005% (w/v) PS20 7.0 ±0.15 10 mM histidine-HCl 10% trehalose  25 mM proline  0.02% (w/v) PS207.0 ± 0.15 10 mM histidine-HCl 10% trehalose  50 mM proline  0.02% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  75 mM proline  0.02%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  25 mM proline 0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose  50 mMproline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10% trehalose 75 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 10%trehalose  50 mM  0.02% (w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol — 0.005% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol —  0.02% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol —  0.05% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol  5 mM 0.005% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl 165 mM glycerol 10 mM 0.005% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol 15 mM 0.005%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol  5mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mMglycerol 10 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol 15 mM  0.02% (w/v) PS20 methionine 7.0 ±0.15 10 mM histidine-HCl 165 mM glycerol  5 mM  0.05% (w/v) PS20methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol 10 mM  0.05%(w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol 15mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 165 mMglycerol (3%) 25 mM proline 0.005% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol (3%) 50 mM proline 0.005% (w/v) PS20 7.0 ±0.15 10 mM histidine-HCl 165 mM glycerol (3%) 75 mM proline 0.005% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol (3%) 25 mM proline 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol (3%) 50mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mMglycerol (3%) 75 mM proline  0.02% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 165 mM glycerol (3%) 25 mM proline  0.05% (w/v) PS20 7.0 ±0.15 10 mM histidine-HCl 165 mM glycerol (3%) 50 mM proline  0.05% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol (3%) 75 mM proline 0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 165 mM glycerol (3%) 50mM  0.02% (w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl 275 mMglycerol (5%) — — 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) —0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) — 0.02% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) — 0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%)  5mM 0.005% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mMglycerol (5%) 10 mM 0.005% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 275 mM glycerol (5%) 15 mM 0.005% (w/v) PS20 methionine7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%)  5 mM  0.02% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) 10mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mMglycerol (5%) 15 mM  0.02% (w/v) PS20 methionine 7.0 ± 0.15 10 mMhistidine-HCl 275 mM glycerol (5%)  5 mM  0.05% (w/v) PS20 methionine7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) 10 mM  0.05% (w/v)PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol (5%) 15mM  0.05% (w/v) PS20 methionine 7.0 ± 0.15 10 mM histidine-HCl 275 mMglycerol 25 mM proline 0.005% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl275 mM glycerol 50 mM proline 0.005% (w/v) PS20 7.0 ± 0.15 10 mMhistidine-HCl 275 mM glycerol 75 mM proline 0.005% (w/v) PS20 7.0 ± 0.1510 mM histidine-HCl 275 mM glycerol 25 mM proline  0.02% (w/v) PS20 7.0± 0.15 10 mM histidine-HCl 275 mM glycerol 50 mM proline  0.02% (w/v)PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol 75 mM proline  0.02%(w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol 25 mM proline 0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol 50 mMproline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mM glycerol75 mM proline  0.05% (w/v) PS20 7.0 ± 0.15 10 mM histidine-HCl 275 mMglycerol 50 mM  0.02% (w/v) PS20 arginine-HCl 6.5 ± 0.15 10 mMhistidine-HCl  5% sucrose 50 mM  0.02% (w/v) PS20 arginine-HCl 6.5 ±0.15 10 mM histidine-HCl  8% sucrose 50 mM  0.02% (w/v) PS20arginine-HCl 6.5 ± 0.15 10 mM histidine-HCl 10% sucrose 50 mM  0.02%(w/v) PS20 arginine-HCl 6.5 ± 0.15 10 mM histidine-HCl 165 mM mannitol50 mM  0.02% (w/v) PS20 arginine-HCl 6.5 ± 0.15 10 mM histidine-HCl 275mM mannitol 50 mM  0.02% (w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mMhistidine-HCl  5% sucrose 50 mM  0.02% (w/v) PS20 arginine-HCl 7.0 ±0.15 10 mM histidine-HCl  8% sucrose 50 mM  0.02% (w/v) PS20arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl — 150 mM   0.02% (w/v) PS20arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl 10% sucrose 50 mM  0.02%(w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl 165 mM mannitol(3%) 50 mM  0.02% (w/v) PS20 arginine-HCl 7.0 ± 0.15 10 mM histidine-HCl275 mM mannitol (5%) 50 mM  0.02% (w/v) PS20 arginine-HCl

TABLE 1C List of sodium phosphate buffer-based formulations pH BufferTonicity agent Excipient Surfactant 6.5 ± 0.15 10 mM sodium  5%sucrose - - - 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5%sucrose  5 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  5% sucrose 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium  5% sucrose 15 mM methionine 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose  5 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose 10 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5%sucrose 15 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  5% sucrose  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  5% sucrose 10 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose 15 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose 25 mM proline0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose 50 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  5%sucrose 25 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 5% sucrose 50 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  5% sucrose 75 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.1510 mM sodium  5% sucrose 25 mM proline 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  5% sucrose 50 mM proline 0.05% (w/v) PS20 phosphate6.5 ± 0.15 10 mM sodium  5% sucrose 75 mM proline 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  5% sucrose 50 mM arginine- 0.02%(w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mM sodium  8% sucrose — 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose — 0.02% (w/v)PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose — 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose  5 mM methionine 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 10 mMmethionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8%sucrose 15 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  8% sucrose  5 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  8% sucrose 10 mM methionine 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 15 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose  5 mMmethionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8%sucrose 10 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  8% sucrose 15 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  8% sucrose 25 mM proline 0.005% (w/v) PS20  phosphate6.5 ± 0.15 10 mM sodium  8% sucrose 50 mM proline 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 75 mM proline 0.005% (w/v)PS20  phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 25 mM proline 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 50 mM proline0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose 75 mMproline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% sucrose25 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8%sucrose 50 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 8% sucrose 75 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  8% sucrose 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 6.5 ±0.15 10 mM sodium 10% sucrose — — phosphate 6.5 ± 0.15 10 mM sodium 10%sucrose — 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10%sucrose — 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose— 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose  5 mMmethionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10%sucrose 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 10% sucrose 15 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 10% sucrose  5 mM methionine 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 10 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 15 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10%sucrose  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 10% sucrose 10 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 10% sucrose 15 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 25 mM proline 0.005% (w/v)PS20  phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 50 mM proline 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 75 mM proline0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose 25 mMproline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% sucrose50 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10%sucrose 75 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium10% sucrose 25 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 10% sucrose 50 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.1510 mM sodium 10% sucrose 75 mM proline 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 10% sucrose 50 mM arginine- 0.02% (w/v) PS20 phosphateHCI 6.5 ± 0.15 10 mM sodium 165 mM mannitol — 0.02% (w/v) PS20 phosphate6.5 ± 0.15 10 mM sodium 165 mM mannitol  5 mM methionine 0.005% (w/v)PS20  phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 10 mM methionine0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 15mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 165mM mannitol  5 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 165 mM mannitol 10 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 165 mM mannitol 15 mM methionine 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol  5 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 10 mMmethionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mMmannitol 15 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 165 mM mannitol 25 mM proline 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 165 mM mannitol 50 mM proline 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 75 mM proline 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 25 mMproline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mMmannitol 50 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 165 mM mannitol 75 mM proline 0.02% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 165 mM mannitol 25 mM proline 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 50 mM proline 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM mannitol 75 mMproline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mMmannitol 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mMsodium 275 mM mannitol — 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 275 mM mannitol  5 mM methionine 0.005% (w/v) PS20  phosphate 6.5± 0.15 10 mM sodium 275 mM mannitol 10 mM methionine 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 15 mM methionine0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol  5mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mMmannitol 10 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 275 mM mannitol 15 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 275 mM mannitol  5 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 10 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 15 mMmethionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mMmannitol 25 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 275 mM mannitol 50 mM proline 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 275 mM mannitol 75 mM proline 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 25 mM proline 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 50 mMproline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mMmannitol 75 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 275 mM mannitol 25 mM proline 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 275 mM mannitol 50 mM proline 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 75 mM proline 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM mannitol 50 mMarginine- 0.02% (w/v) PS20 phosphate HCI 7.0 ± 0.15 10 mM sodium  8%sucrose — 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium  5% sucrose— 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium  5% sucrose 10 mMmethionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium  5%sucrose 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 5% sucrose 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 7.0 ± 0.15 10mM sodium  8% sucrose 10 mM methionine 0.02% (w/v) PS20 phosphate 7.0 ±0.15 10 mM sodium  8% sucrose 50 mM proline 0.02% (w/v) PS20 phosphate7.0 ± 0.15 10 mM sodium  8% sucrose 50 mM arginine- 0.02% (w/v) PS20phosphate HCI 7.0 ± 0.15 10 mM sodium 10% sucrose — — phosphate 7.0 ±0.15 10 mM sodium 10% sucrose — 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10mM sodium 10% sucrose — — phosphate 7.0 ± 0.15 10 mM sodium 10% sucrose— 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 10% sucrose 10 mMmethionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 10%sucrose 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium10% sucrose 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 7.0 ± 0.15 10mM sodium 165 mM mannitol — 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium 165 mM mannitol 10 mM methionine 0.02% (w/v) PS20 phosphate 7.0 ±0.15 10 mM sodium 165 mM mannitol 50 mM proline 0.02% (w/v) PS20phosphate 7.0 ± 0.15 10 mM sodium 165 mM mannitol 50 mM arginine- 0.02%(w/v) PS20 phosphate HCI 7.0 ± 0.15 10 mM sodium 275 mM mannitol — 0.02%(w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 275 mM mannitol 10 mMmethionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 275 mMmannitol 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium 275 mM mannitol 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI

TABLE 1D List of sodium phosphate buffer-based formulations pH BufferTonicity agent Excipient Surfactant 6.5 ± 0.15 10 mM sodium  5%trehalose — 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5%trehalose  5 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  5% trehalose 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium  5% trehalose 15 mM methionine 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose  5 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 10 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5%trehalose 15 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  5% trehalose  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  5% trehalose 10 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 15 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 25 mM proline0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 50 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  5%trehalose 75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  5% trehalose 25 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.1510 mM sodium  5% trehalose 50 mM proline 0.02% (w/v) PS20 phosphate 6.5± 0.15 10 mM sodium  5% trehalose 75 mM proline 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 25 mM proline 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 50 mM proline0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose 75 mMproline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  5% trehalose50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mM sodium 8% trehalose — 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8%trehalose — 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8%trehalose — 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8%trehalose  5 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  8% trehalose 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium  8% trehalose 15 mM methionine 0.005% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose  5 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 10 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8%trehalose 15 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium  8% trehalose  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium  8% trehalose 10 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 15 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 25 mM proline0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 50 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium  8%trehalose 75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium  8% trehalose 25 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.1510 mM sodium  8% trehalose 50 mM proline 0.02% (w/v) PS20 phosphate 6.5± 0.15 10 mM sodium  8% trehalose 75 mM proline 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 25 mM proline 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 50 mM proline0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose 75 mMproline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium  8% trehalose50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mM sodium10% trehalose — — phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose —0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose —0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose — 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose  5 mMmethionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10%trehalose 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 10% trehalose 15 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 10% trehalose  5 mM methionine 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 10 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 15 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10%trehalose  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 10% trehalose 10 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 10% trehalose 15 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 25 mM proline 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 50 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 10%trehalose 75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 10% trehalose 25 mM proline 0.02% (w/v) PS20 phosphate 6.5 ± 0.1510 mM sodium 10% trehalose 50 mM proline 0.02% (w/v) PS20 phosphate 6.5± 0.15 10 mM sodium 10% trehalose 75 mM proline 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 25 mM proline 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 50 mM proline0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose 75 mMproline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 10% trehalose50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mM sodium165 mM glycerol — 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165mM glycerol  5 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10mM sodium 165 mM glycerol 10 mM methionine 0.005% (w/v) PS20  phosphate6.5 ± 0.15 10 mM sodium 165 mM glycerol 15 mM methionine 0.005% (w/v)PS20  phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol  5 mM methionine0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 10 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mMglycerol 15 mM methionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 165 mM glycerol  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 165 mM glycerol 10 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 15 mM methionine 0.05%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 25 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 165 mMglycerol 50 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 165 mM glycerol 75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 165 mM glycerol 25 mM proline 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 50 mM proline 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 75 mMproline 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 165 mMglycerol 25 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 165 mM glycerol 50 mM proline 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 165 mM glycerol 75 mM proline 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 165 mM glycerol 50 mM arginine- 0.02%(w/v) PS20 phosphate HCI 6.5 ± 0.15 10 mM sodium 275 mM glycerol — 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol  5 mMmethionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 275 mMglycerol 10 mM methionine 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 275 mM glycerol 15 mM methionine 0.005% (w/v) PS20  phosphate 6.5± 0.15 10 mM sodium 275 mM glycerol  5 mM methionine 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 10 mM methionine 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 15 mMmethionine 0.02% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mMglycerol  5 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 275 mM glycerol 10 mM methionine 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 275 mM glycerol 15 mM methionine 0.05% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 25 mM proline 0.005%(w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 50 mMproline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mM sodium 275 mMglycerol 75 mM proline 0.005% (w/v) PS20  phosphate 6.5 ± 0.15 10 mMsodium 275 mM glycerol 25 mM proline 0.005% (w/v) PS20  phosphate 6.5 ±0.15 10 mM sodium 275 mM glycerol 50 mM proline 0.02% (w/v) PS20phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 75 mM proline 0.02%(w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mM glycerol 25 mMproline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mM sodium 275 mMglycerol 50 mM proline 0.05% (w/v) PS20 phosphate 6.5 ± 0.15 10 mMsodium 275 mM glycerol 75 mM proline 0.05% (w/v) PS20 phosphate 6.5 ±0.15 10 mM sodium 275 mM glycerol 50 mM arginine- 0.02% (w/v) PS20phosphate HCI 7.0 ± 0.15 10 mM sodium  8% trehalose — 0.02% (w/v) PS20phosphate 7.0 ± 0.15 10 mM sodium  5% trehalose — 0.02% (w/v) PS20phosphate 7.0 ± 0.15 10 mM sodium  5% trehalose 10 mM methionine 0.02%(w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium  5% trehalose 50 mM proline0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium  5% trehalose 50 mMarginine- 0.02% (w/v) PS20 phosphate HCI 7.0 ± 0.15 10 mM sodium  8%trehalose 10 mM methionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium  8% trehalose 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ± 0.1510 mM sodium  8% trehalose 50 mM arginine- 0.02% (w/v) PS20 phosphateHCI 7.0 ± 0.15 10 mM sodium 10% trehalose — — phosphate 7.0 ± 0.15 10 mMsodium 10% trehalose — 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium 10% trehalose — — phosphate 7.0 ± 0.15 10 mM sodium 10% trehalose— 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 10% trehalose 10 mMmethionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 10%trehalose 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium 10% trehalose 50 mM arginine- 0.02% (w/v) PS20 phosphate HCI 7.0± 0.15 10 mM sodium 165 mM glycerol — 0.02% (w/v) PS20 phosphate 7.0 ±0.15 10 mM sodium 165 mM glycerol 10 mM methionine 0.02% (w/v) PS20phosphate 7.0 ± 0.15 10 mM sodium 165 mM glycerol 50 mM proline 0.02%(w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 165 mM glycerol 50 mMarginine- 0.02% (w/v) PS20 phosphate HCI 7.0 ± 0.15 10 mM sodium 275 mMglycerol — 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mM sodium 275 mMglycerol 10 mM methionine 0.02% (w/v) PS20 phosphate 7.0 ± 0.15 10 mMsodium 275 mM glycerol 50 mM proline 0.02% (w/v) PS20 phosphate 7.0 ±0.15 10 mM sodium 275 mM glycerol 50 mM arginine- 0.02% (w/v) PS20phosphate HCI

TABLE lE List of various formulations pH Buffer Tonicity agent ExcipientSurfactant 5.0 ± 0.15 10 mM sodium acetate 2.5% glycerol None None 5.0 ±0.15 10 mM sodium citrate 10% trehalose None None 5.5 ± 0.15 10 mMsodium citrate 0.9% NaCl None None 6.0 ± 0.15 10 mM sodium citrate 10%trehalose None None 6.5 ± 0.15 10 mM sodium citrate 10% sucrose NoneNone 5.5 ± 0.15 10 mM histidine-HCI 10% trehalose None None 6.0 ± 0.1510 mM histidine-HCI 10% trehalose None None 7.0 ± 0.15 10 mMhistidine-HCI 275 mM glycerol None None (2.5%) 7.0 ± 0.15 10 mMhistidine-HCI 0.9% NaCl None None 6.5 ± 0.15 10 mM sodium 275 mMglycerol None 0.02% (w/v) PS20 phosphate (2.5%) 6.5 ± 0.15 10 mM sodium150 mM NaCl None 0.02% (w/v) PS20 phosphate (0.9%) 6.5 ± 0.15 10 mMsodium 50 mM NaCl 100 mM arginine- 0.02% (w/v) PS20 phosphate (0.3%) HCI6.5 ± 0.15 10 mM sodium 150 mM NaCl None None phosphate (0.9%) 6.5 ±0.15 10 mM sodium 130 mM NaCl 50 mM proline 0.02% (w/v) PS20 phosphate(0.7%) 6.5 ± 0.15 10 mM sodium 150 mM NaCl 10 mM methionine 0.02% (w/v)PS20 phosphate (0.9%) 6.5 ± 0.15 10 mM histidine-HCI 10% sucrose — — 7.0± 0.15 10 mM histidine-HCI 10% sucrose — —

Analytical methods were developed and verified for subsequent testing ofthe physicochemical characteristics of different formulation candidatesunder stress conditions. The analytical methods used are outlined inTable 2.

TABLE 2 Analytical methods for protein stability and liquid productcharacterization. Method Purpose Reversed-phase ultra-performancePurity: chemical changes, content liquid chromatography (RP-UPLC) Highperformance size-exclusion Purity: monomer, higher molecularchromatography (HP-SEC) weight species UV spectroscopy Peptide content,turbidity Micro-Flow Imaging (MFI) Quantification and visualization ofsubvisible particles (~1-100 μm) Visual inspection Visible particlesaccording to Ph. Eur pH pH Osmolality Osmolality

The stress conditions applied to the different formulations arepresented in Table 3. Some formulations were subjected to acceleratedstorage for 4 weeks at 25° C. and 40° C. while others were subjected toaccelerated storage for at least 3 months at 5° C., 25° C. and 40° C.

TABLE 3 Stress conditions for formulations. Condition Parameters Timepoints Unstressed Formulation T0 Mechanical Mechanical stress: T-mechstress shaking Freeze-thawing Five freeze-thaw cycles T-FT stressStorage at 5° C. Storage at 2-8° C. T-1m_5° C. T-3m_5° C. Storage at 25°Storage at 25° C. and T-1m_25° C. C. 60% r.h. T-3m_25° C. Storage at 40°Storage at 40° C. and T-1m_40° C. C. 75% r.h. T-3m_40° C.

Each of the conditions outlined at different time points in Table 3 maybe considered as a stress condition (except when unstressed).

Example 2: Stability Testing

Formulations of Table 4 were prepared and tested in one or more of thefollowing sets of experiments.

Visual Inspection

The vials are inspected for the presence or absence of visible particlesunder gentle, manual, radial agitation for 5 seconds in front of a whitebackground and for 5 seconds in front of a black background according tothe European Pharmacopoeia (8th edition; monograph 2.9.20).

Based on the above, it was determined that at T0, substantial visibleparticle formation (precipitation) was detected in one formulation at pH5.0±0.15, containing 10 mM sodium citrate and 10% trehalose and inanother formulation at pH 5.5±0.15, containing 10 mM histidine-HCl and10% trehalose. At T0, the remaining twenty-six formulations testedshowed no or very low amounts of visible particles.

No substantial impact on the visible particle content was observed uponstorage for 1 month at 5° C. in formulations tested under theseconditions.

After storage for 1 month at 25° C. and/or 1 month at 40° C., highamount of visible particles was detected in formulations containingsodium acetate, sodium citrate or histidine-HCl at low pH (5.5±0.15) andin some formulations at pH of between 6.0±0.15-7.0±0.15 containinghistidine-HCl and 10% trehalose or sucrose (without excipient orsurfactant).

Despite the absence of surfactants, other formulations containinghistidine or phosphate buffer and sugar performed well with respect tothe determined visible particle contents after storage for 1 month at25° C. and at 40° C.

Very few visible particles were detected in tested formulations at pH7.0±0.15 containing histidine-HCl, a tonicity agent, a surfactant withor without amino acid excipients or at pH 6.5±0.15 containing sodiumphosphate, a tonicity agent with or without a surfactant when stored forthree months at 5° C., 25° C. and/or 40° C.

Freeze-thaw stress led to substantial formation of visible particlesonly in two histidine-based formulations containing arginine(formulations at pH 7.0±0.15 each containing either 10 mM histidine-HCl,50 mM arginine-HCl, 3% mannitol and 0.02% (w/v) PS20 or 10 mMhistidine-HCl, 150 mM arginine-HCl and 0.02% (w/v) PS20). Additionally,phase separation was observed in most of the tested formulations, whichdisappeared after gentle homogenization of the samples.

Mechanical stress led to a strong increase in visible particle contentfor some of the polysorbate-free formulations tested.

pH

pH is measured with a calibrated pH meter.

The pH of various formulations is measured at room temperature (e.g.,generally between 20 and 25° C. and more particularly between 22-24°C.). The pH values of the tested formulations remained relativelyconstant throughout the stability study. Generally, the histidine-basedformulations showed more fluctuations than the phosphate-basedformulations.

Osmolality

At T0, the osmolality values for formulations that contain the peptideof interest, a buffering agent and a tonicity agent were in rangebetween 299-369 mOsmol/kg. The values for formulations that also containan excipient and/or a surfactant were between 276-313. A lowerosmolality value of 235 mOsmol/kg was measured for one of theformulations containing proline. If desired, the osmolality of suchformulations could be raised by increasing the sugar content.

UV Spectroscopy

UV measurements are performed in 96-well plates. Absorbance (A) ismeasured at 280 nm (A280 nm) and 320 nm (A320 nm).

The peptide concentrations at T0 and/or after storage for 1 month at 25°C. were within the target range (0.3±0.05 mg/ml) for all formulationsexcept for formulations that showed visible precipitation, in some ofthe citrate-based formulations and/or those at pH of 5.5±0.01 or lower.Storage for 1 month at 40° C. had no effect on the peptide content; thepeptide concentration did not vary or remained well within target range,except for one of the precipitated formulations.

Some of the polysorbate-free formulations tested were negativelyimpacted after mechanical stress.

Freeze-thaw stress as well as storage at 5° C., for 1 month had noinfluence on the peptide content in the tested formulations. The peptidecontent also remained stable for the formulations stored for threemonths at 5° C. and 25° C.

After storage at 40° C. for 3 months, a slight decrease in peptideconcentration was observed in the tested formulations. However, they allremained well within target range.

Turbidity

The optical density (OD) values determined at 350 nm (OD350 nm) andeither 550 nm (OD550 nm) or 580 nm (OD580 nm) are used to assess theturbidity of the samples.

At T0, turbidity (OD350 nm and OD550 nm) was very low for allformulations except for one of the formulations that had shownprecipitation during preparation.

Turbidity values was very low after freeze-thaw stress and mechanicalstress in tested formulations, and after storage for 1 month (alltemperatures). However, the polysorbate-free formulations tested aftermechanical stress showed very high turbidity (OD350 nm>0.10). Uponstorage at 25° C. for 1 month, the turbidity values remained low. Onlyupon storage at 40° C. for 1 month, the turbidity (OD350 nm) increasedslightly for some of the formulations including those that showedprecipitation at T0. The OD550 nm values did not change substantiallyupon storage at 25° C. and 40° C.

After storage for 3 months, a slight increase in turbidity was alsoobserved for some of the formulations stored at 40° C. However, withOD350 nm values of 0.03 and 0.02 the turbidity of these samples is stillgenerally low.

Micro-Flow Imaging™ (MFI)

Flow imaging microscopy allows enumerating and visualization ofsubvisible particles present in a sample.

In this technique bright-field images are captured in successive framesas a continuous sample stream passes through a flow cell positioned inthe field of view of a microscopic system. The digital images of theparticles present in the sample are processed by image morphologyanalysis software that allows their quantification in size and count(Zölls, S et al., The AAPS Journal. Vol 15(4), p. 1200, 2013).

Micro-Flow Imaging measurements are conducted on a 5200particle-analyzer system.

MFI View System Software (MVSS) version 2-R2-6.1.20.1915 is used toperform the measurements and MFI View Analysis Suite (MVAS) softwareversion 1.3.0.1007 is used to analyze the samples.

At T0, MFI revealed low concentrations of subvisible particles in allformulations except in formulations where precipitation was observed(i.e., high concentration of subvisible particles was measured)

Storage for 1 month at 5° C. had no impact on the subvisible particleconcentrations in any of the formulations tested.

Storage at 25° C. or 40° C. for 1 month revealed good stability of mostformulations, since only slight increases in subvisible particle countswere observed yet the concentration of subvisible particles remainedlow.

At T−1m_25° C. and/or T−1m_40° C., a high concentration of subvisibleparticles was also observed for one of the polysorbate-free formulation.

At T−1m_25° C. and/or T−1m_40° C., a high concentration of subvisibleparticles was also observed for other formulations such as those thatshowed precipitation at T0, those at pH of 5.5±0.01 or below and oneformulation at pH 6.0±0.01 containing 10 mM histidine-HCl and 10%trehalose showed a substantial change in subvisible particle content.

Low concentration of subvisible particles and very good stability wasobserved for the formulations tested after storage at 5° C., 25° C. and40° C. for 3 months.

Freeze-thaw stress had no impact on the particle counts for most of theformulations tested under those conditions. Only the two histidine-basedformulations containing arginine showed a clear increase in subvisibleparticles under these conditions which was consistent with visualinspection.

Most formulations showed a very good stability towards mechanicalstress. Only the two polysorbate-free formulations tested showed asignificant increase in subvisible particle content which was consistentwith visual inspection.

Generally, citrate formulations and/or those having a pH of 5.0-5.5appeared to have the greatest number of subvisible particles whereasformulations containing histidine or phosphate appear to have lowernumber of subvisible particles.

High Performance Size-Exclusion Chromatography (HP-SEC)

In unstressed ROSE-010 samples, a clear monomer peak as well as peakscorresponding to high molecular weight species (HMWS) and low molecularweight species (LMWS) were observed at 280 nm. The ability of the HP-SECmethod to detect changes in the monomer, HMWS and LMWS content wastested with stressed samples. The method was capable of detectingdegradation products, as indicated by a slight increase of the HMWScontent after one week storage at 40° C. Multiple freeze-thaw cycles hadno influence on the SEC profile. Mechanical stress led to a substantialloss of the overall signal intensity.

Relative Monomer Content

At T0, the monomer content of several of the tested formulations wasabout 96%.

However, at T0, the citrate formulations showed a lower monomer contentof about 80%. The monomer content of formulations containing NaCl astonicity agent was also low (ranging from 67.6% to 81.9%). The monomercontent of the two histidine-based formulations containing arginine wasalso very low at T0 (64.9%). After storage for 1 month at 25° C., themonomer content decreased even more for these formulations (monomercontent ranging from 63.2 to 70.7% for citrate-based formulations andfrom 44.4 to 59.6% for formulations containing NaCl). The monomercontent generally decreased even further when formulations were storedfor 1 month at 40° C. (especially those containing NaCl). The monomercontent tested for formulations containing NaCl also decreased uponstorage for 1 month at 5° C. compared to T0.

After storage for 1 month at 5° C. or for 1 month at 25° C., the monomercontent of the remaining formulations tested was relatively unchanged,with the exception of one the two histidine-based formulationscontaining arginine (66.5% at 5° C. and 78.3% at 25° C.). The monomercontent of these formulations decreased slightly under storage for 1month at 40° C.

Monomer content of 96% and higher at T−1m_5° C., of 95.8% or higher atT−1m_25° C. and of 94.3% or higher at T−1 m_40° C. were measured in themost stable formulations.

Storage for 3 months at 5° C. had no effect on the monomer content forthe formulations tested under these conditions (monomer content of 95.9%or higher). Storage at 25° C. for 3 months led to a slight decrease inthe monomer content for two of these formulations (decrease of 0.7% and1.6% respectively compared to T0, i.e., monomer content of 95.1 or 95.3%or higher). Storage at 40° C. for 3 months led to a slight decrease inmonomer content for three of the formulations tested and to a modestdecrease for two of these formulations (decrease of 7.2% and 8.2%compared to T0). Three formulations still had a monomer content of 93.0%or higher at T−3m_40° C.

In comparison with T0 unstressed sample, mechanical stress led to asubstantial decrease in monomer content in two of the polysorbate-freeformulations tested (with approximately 54-60% decrease). All otherformulations remained relatively stable upon mechanical stress.

Freeze-thaw stress had no substantial impact on the monomer content inany of the formulations tested under these conditions.

Relative Content of HMWS

At T0, the relative aggregate content was about 1% for mosthistidine-based formulations. The citrate-based formulation showed ahigher aggregate content of about 15 to 17.7% (at 280 nm). Forcitrate-based formulation, the aggregate content further increased withboth heat stress conditions (ranging from about 27 to 33.7% at T−1m_25°C. and from about 21.7 to 38.6% at T−1m_40° C., 280 nm).

The relative aggregate content for most of the histidine-basedformulations stayed very low (ranging from 0.8% to 1.6%) at T−1m_5° C.and/or T−1m_25° C. and increased slightly at T−1m_40° C. (ranging from1.2% to 3.0%). The two histidine-based formulations containing arginineshowed a relative aggregate content of between 10.2 to 19.7% at 5° C.,10.3 to 20.0% at 25° C. and 13.1 to 20.3% at 40° C. One formulation atpH 7.0±0.15 containing histidine-HCl and NaCl showed a substantialincrease in aggregate content after storage for 1 month at 40° C. (about30% at T−1m_40° C., 280 nm). A minor increase was observed for otherformulations (ranging from about 1.2 to 3.0% at T−1m_40° C., 280 nm).

After mechanical stress, a clear increase in the relative HMWS contentwas observed for one of the polysorbate-fee formulations (reaching11.1%).

Freeze-thaw stress had no substantial effect on the relative HMWScontent in of the formulations tested under these conditions.

For some of the formulations, especially those containing sodiumphosphate as buffering agent, a different peak shape, which wasinterpreted as high-molecular-weight species, was already present at T0.However, for two of such formulations (one containing glycerol andpolysorbate and the other containing sucrose and polysorbate) a very lowaggregate content at T0, T−1m_5° C. and T−1m_25° C. (1.6% or lower) wasmeasured and increased slightly at T−1m_40° C. (reaching 4.9% and 2.6%respectively).

Storage for 3 months at 5° C. had no influence on the relative HMWScontent in any of the formulations tested. Storage for 3 months at 25°C. led to a slight increase in the HMWS content in three of theseformulations (reaching 1.7%, 1.9% and 2.2%). Storage for 3 months at 40°C. led to a substantial increase in HMWS content (from 2.2 to 8.2%) forall formulations tested, which was strongest for two of theseformulations (5.7% and 8.2%). Still the HMWS content was below 10% forall formulations.

Relative Content of LMWS

At T0, the relative amount of LMWS was about 2-3.3% at 280 nm for allformulations tested.

Mechanical stress led to a substantial increase in the relative LMWScontent, especially for the two polysorbate-free formulations(49.5-50.1%), but also for one of the two histidine-based formulationcontaining arginine (15.0%) and in several formulations containing NaClas tonicity agent (ranging from 16.5 to 22.8%). These formulations alsoshowed a high LMWS content upon storage for 1 month at 5° C., 25° C. and40° C. For example, the two histidine-based formulations containingarginine had a LMWS content after storage at 5° C. for 1 month rangingfrom 13.7% to 34.8%.

Freeze-thaw stress led to a clear increase in the relative LMWS contentfor formulations containing NaCl as tonicity agent (from 8.9 to 18.1%);the LMWS content for all other formulations tested remained unchanged.

Storage for 1 month at 5° C., 25° C. and 40° C. had no substantialimpact on the relative LMWS content of the remaining formulations(reaching 1.6 to 5.5%).

Storage for 3 months at 5° C. or 25° C. had no impact on the relativeLMWS content for any of the formulations tested. Storage for 3 months at40° C. led to a small increase in the relative LMWS content for alltested formulations, which reached 4.6 to 5.1% for two of theseformulations.

The total peak remained stable amongst all time points and samples. Theonly exceptions were the polysorbate-free formulations after mechanicalstress, which showed almost a complete loss of signal.

Reversed Phase Ultra-Performance Liquid Chromatography (RP-UPLC)

The stability-indicating potential of the RP-UPLC method was tested withstressed samples. The RP-UPLC method is able to detect degradationproducts, as indicated by new peaks arising after one-week storage at40° C. Precipitation observed in samples after mechanical stress led toa decrease of the total peak area in RP-UPLC chromatogram.

The RP-UPLC is sensitive for the detection of impurities and can be usedfor the analysis of the formulations.

Main Peak Content

At T0, the relative main peak area of the tested formulations rangedbetween 94.7% and 97.1%.

Storage at 5° C. for 1 month had no impact on the relative main peakarea in any of formulations tested.

After storage at 25° C. for 1 month, the relative main peak area wasslightly lower for the formulations that showed precipitation atpreparation (91.6% and 93.1%) but generally remained in the same rangesfor the other formulations (from 93.1 to 96.3%).

For some formulations, storage at 40° C. for 1 month further decreasedthe main peak content (from 83.3% to 89.2%). For example, a formulationat pH 5.0 that contains sodium citrate and trehalose showed a main peakcontent of 83.3%, one formulation at pH 6.0 containing sodium citrateand trehalose showed a main peak content of 89.2%, one formulation at pH7.0 containing histidine-HCl and sucrose and one formulation at pH 7.5containing sodium phosphate and sucrose, both without excipients andsurfactants showed a main peak content of 89.0% and 87.6% respectively.Generally, the main peak content remained in the range of between 90.0%to 94.5% for the other formulations tested.

Storage at 5° C. for 3 months had no effect on the relative main peakarea in any of the formulations tested. Storage at 25° C. for 3 monthsled to a small decrease in the relative main peak area (varying from94.2 to 94.9%), while storage at 40° C. for 3 months led to asubstantial decrease, with the lowest relative main peak area observedof 82.0-83.0%).

Freeze-thaw stress had no impact on the relative main peak area in anyof formulations tested under these conditions.

Mechanical stress had no substantial impact on most of formulationstested under these conditions, except for the polysorbate-freeformulations (relative main peak area of 72.0% and 89.8%).

Total Impurity Content

The inverse trend was observed for the total impurity content. The totalimpurity content of most formulations at T0 varied from 2.9 to 3.9%. Thehighest level of impurity at T0 was measured for one of the formulationsthat showed precipitation during preparation (5.3%).

An increase in total impurities was observed for the polysorbate-freeformulations (10.3% and 28.0% total impurities) after mechanical stress.There was no significant impact on total impurity content forformulations submitted to freeze-thaw.

The level of impurities was slightly higher after storage at 25° C. for1 month generally ranging from 4.2% to 5.5% with the exception of theformulations that showed precipitation during preparation (6.9% and8.4%).

Storage at 40° C. for 1 month further increased the level of impuritiesin all formulations. The four formulations that showed a decrease in themain peak (from 83.3% to 89.2%) also showed an increase in the level ofimpurities (from 10.8 to 16.7%). However, for most of thehistidine-based or sodium phosphate-based formulations tested, the levelof impurities remained below 10%.

No impact of storage at 5° C. was detected even after 3 or 6 months inthe formulations tested. A gradual increase for formulations stored at25 and 40° C. for 3 months was observed. The total impurity content atT3m-5° C. was between 3.5% to 4.0%, the total impurity content atT3m-25° C. was between 5.1% to 5.8% and the total impurity content atT3m-40° C. was between 11.7% to 18.0%.

The total peak area remained constant for most time points in theformulations tested. For the T-mech samples and the T−1m_40° C. samples,slightly higher peak areas were observed. For the polysorbate-freeformulations after mechanical stress, an almost complete loss of signalwas observed (data not shown).

Pharmaceutical formulations appear more stable at a higher pH (e.g., pH6.0±0.15 to 7.5±0.15) and as such formulations containing low pH buffersappear less stable. A pH of between 6.5±0.15 to 7.0±0.15 appearsbeneficial for peptide stability. Surfactants and amino acid stabilizersmay help in preserving the stability of the formulations especiallyagainst stress conditions.

Formulations comprising GLP-1 or other GLP-1 analogues are also expectedto be stable under the same conditions described herein and morespecifically in the experimental section.

TABLE 4 Composition   F1: 10 mM sodium acetate pH 5.0; 2.5% glycerol F2:10 mM sodium citrate pH 5.0; 10% trehalose F3: 10 mM sodium citrate pH5.5; 0.9% NaCl F4: 10 mM sodium citrate pH 6.0; 10% trehalose F5: 10 mMsodium citrate pH 6.5; 10% sucrose F6: 10 mM histidine-HCl pH 5.5; 10%trehalose F7: 10 mM histidine-HCl pH 6.0; 10% trehalose F8: 10 mMhistidine-HCl pH 6.5; 10% sucrose F9: 10 mM histidine-HCl pH 7.0; 10%sucrose F10: 10 mM histidine-HCl pH 7.0; 5% mannitol F11: 10 mMhistidine-HCl pH 7.0; 0.9% NaCl F12: 10 mM sodium phosphate pH 6.5; 10%sucrose F13: 10 mM sodium phosphate pH 7.0; 10% sucrose F14: 10 mMsodium phosphate pH 7.5; 10% sucrose F15; 10 mM histidine-HCl pH 7.0;165 mM mannitol; 50 mM arginine-HCl, 0.02% (w/v) PS20 F16: 10 mMhistidine-HCl pH 7.0; 165 mM mannitol; 50 mM proline; 0.02% (w/v) PS20F17: 10 mM histidine-HCl pH 7.0; 275 mM mannitol; 10 mM methionine;0.02% (w/v) PS20 F18: 10 mM histidine-HCl pH 7.0; 275 mM mannitol; 0.02%(w/v) PS20 F19: 10 mM histidine-HCl pH 7.0; 150 mM arginine-HCl; 0.02%(w/v) PS20 F20: 10 mM histidine-HCl pH 7.0; 275 mM glycerol F21: 10 mMphosphate-Na pH 6.5; 275 mM glycerol; 0.02% (w/v) PS20 F22: 10 mMphosphate-Na pH 6.5; 150 mM NaCl; 0.02% (w/v) PS20 F23: 10 mMphosphate-Na pH 6.5; 50 mM NaCl; 100 mM arginine-HCl; 0.02% (w/v) PS20F24: 10 mM phosphate-Na pH 6.5; 150 mM NaCl F25: 10 mM phosphate-Na pH6.5; 130 mM NaCl; 50 mM proline; 0.02% (w/v) PS20 F26: 10 mMphosphate-Na pH 6.5; 150 mM NaCl; 10 mM methionine; 0.02% (w/v) PS20F27: 10 mM phosphate-Na pH 6.5; 8% sucrose; 0.02% (w/v) PS20 F28: 10 mMhistidine-HCl pH 7.0; 8% sucrose; 10 mM methionine; 0.02% (w/v) PS20

The aspects, embodiments and examples described herein are illustrativeand are not meant to be limitative. Variations of the foregoingembodiments, including alternatives, modifications and equivalents, areintended by the inventors to be encompassed by the present disclosure.Citations listed in the present application are incorporated herein byreference.

SEQUENCE LISTING

SEQ ID NO: 1 X₁X₂EGTFTSDVSSYLX₃GQAAKX₄FIAWLVKGRX₅Wherein X₁ may be L-histidine, D-histidine, desaminohistidine,2-amino-histidine, beta-hydroxy-histidine, homohistidine,alpha-fluoromethylhistidine or alpha-methyl-histidineWherein X₂ may be V, A, G, T, I or alpha-methyl-Ala

Wherein X₃ may be E, Q, A, T, S or G Wherein X₄ may be E, Q, A, T, S orG

Wherein X₅ may be absent, Gly-NH₂ or Gly-OH

(GLP-1 (7-37) SEQ ID NO: 2 H-HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG-OH(ROSE-010) SEQ ID NO: 3 H-HVEGTFTSDVSSYLEGQAAKEFIAWLVKGRG-OH(SEQ ID NO: 2 without terminal glycine residue) SEQ ID NO: 4H-HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR-NH₂(ROSE-010 without terminal glycine residue) SEQ ID NO: 5H-HVEGTFTSDVSSYLEGQAAKEFIAWLVKGR-NH₂

STATEMENTS OF INVENTION

-   -   1. A liquid pharmaceutical formulation comprising glucagon-like        peptide 1 (GLP-1) or a GLP-1 analogue thereof as defined in SEQ        ID NO:1, a buffering agent, a tonicity agent, and optionally an        excipient and/or surfactant.    -   2. The pharmaceutical formulation of statement 1, wherein the        pharmaceutical formulation is at a pH of between 6.5±0.5 to        7.5±0.5.    -   3. The pharmaceutical formulation of statement 1, wherein the        pharmaceutical formulation is at a pH of between 6.0±0.15 to        7.5±0.15.    -   4. The pharmaceutical formulation of statement 1 or 2, wherein        the GLP-1 or analogue thereof is GLP-1 (7-37) and comprises the        amino acid sequence set forth in SEQ ID NO:2.    -   5. The pharmaceutical formulation of any one of statements 1 to        4, wherein the GLP-1 or analogue thereof is ROSE-010 and        comprises the amino acid sequence set forth in SEQ ID NO:3.    -   6. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 10 μg/ml to 2 mg/ml.    -   7. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 50 μg/ml to 2 mg/ml.    -   8. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 100 μg/ml to 1 mg/ml.    -   9. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 10 μg/ml to 1 mg/ml.    -   10. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 50 μg/ml to 750 μg/ml.    -   11. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 100 μg/ml to 500 μg/ml.    -   12. The pharmaceutical formulation of any one of statements 1 to        5, wherein the GLP-1 or analogue thereof is at a concentration        of between 200 μg/ml to 500 μg/ml.    -   13. The pharmaceutical formulation of any one of statements 1 to        12, wherein the pharmaceutical formulation is at a pH of between        6.5±0.15 to 7.0±0.15.    -   14. The pharmaceutical formulation of any one of statements 1 to        13, wherein the pharmaceutical formulation comprises a        surfactant.    -   15. The pharmaceutical formulation of statement 14, wherein the        surfactant is a non-ionic surfactant.    -   16. The pharmaceutical formulation of statement 14, wherein the        surfactant is polysorbate 20, polysorbate 40, polysorbate 60,        polysorbate 65, polysorbate 80, Triton X-100, poloxamer,        pluronic F-68 or combination thereof.    -   17. The pharmaceutical formulation of any one of statements 1 to        13, wherein the pharmaceutical formulation comprises        polysorbate.    -   18. The pharmaceutical formulation of any one of statements 1 to        17, wherein the buffering agent is acetate, carbonate, citrate,        histidine, maleate, phosphate, succinate, tartrate,        tromethamine.    -   19. The pharmaceutical formulation of any one of statements 1 to        18, wherein the tonicity agent is a sugar.    -   20. The pharmaceutical formulation of any one of statements 1 to        18, wherein the tonicity agent is selected from the group        consisting of dextrose, glucose, glycerin glycerol, mannitol,        potassium chloride, sodium chloride, sodium sulfate, sorbitol,        sucrose, trehalose or combination thereof.    -   21. The pharmaceutical formulation of any one of statements 1 to        20, wherein the pharmaceutical formulation comprises an        excipient.    -   22. The pharmaceutical formulation of statement 21, wherein the        excipient comprises an amino acid excipient.    -   23. The pharmaceutical formulation of statement 22, wherein the        amino acid excipient is alanine, arginine, isoleucine,        glutamate, glycine, leucine, methionine, proline, and the like        and combination thereof    -   24. The pharmaceutical formulation of any one of statements 1 to        23, wherein the pharmaceutical formulation is at a pH of        6.5±0.5.    -   25. The pharmaceutical formulation of any one of statements 1 to        23, wherein the pharmaceutical formulation is at a pH of        7.0±0.5.    -   26. The pharmaceutical formulation of any one of statements 1 to        23, wherein the pharmaceutical formulation is at a pH of        7.5±0.5.    -   27. The pharmaceutical formulation of any one of statements 1 to        23, wherein the formulation is at pH 7.0±0.15.    -   28. The pharmaceutical formulation of any one of statements 1 to        23, wherein the formulation is at pH 6.5±0.15.    -   29. The pharmaceutical formulation of statement 1 or 28, wherein        the GLP-1 analogue is as set forth in SEQ ID NO:3 and is at a        concentration of from about 50 μg/ml to about 2 mg/ml.    -   30. The pharmaceutical formulation of statement 1 or 28, wherein        the GLP-1 analogue is as set forth in SEQ ID NO:3 and is at a        concentration of from about 100 μg/ml to about 1 mg/ml.    -   31. The pharmaceutical formulation of statement 1 or 28, wherein        the GLP-1 analogue is as set forth in SEQ ID NO:3 and is at a        concentration of from about 100 μg/ml to about 500 μg/ml.    -   32. The pharmaceutical formulation of statement 1 or 31, wherein        the GLP-1 analogue is as set forth in SEQ ID NO:3 and is at a        concentration of 300 μg/ml±20%.    -   33. A pharmaceutical formulation as described in the present        disclosure.    -   34. A pharmaceutical formulation as described in Table 1A, Table        1B, Table 10, Table 1D, Table 1E or Table 4.    -   35. The pharmaceutical formulation of any one of statements 1 to        34, in a unit-dose vial, multi-dose vial, cartridge or        pre-filled syringe.    -   36. The pharmaceutical formulation of any one of statements 1 to        34, wherein the formulation is encapsulated.    -   37. A single-dose or multiple-dose container comprising the        pharmaceutical formulation of any one of statements 1 to 34.    -   38. A pre-filled syringe comprising the pharmaceutical        formulation of any one of statements 1 to 34.    -   39. A method of treating a disorder or condition in which        administration of a GLP-1 or a GLP-1 analogue is indicated, the        method comprising administering the pharmaceutical formulation        of any one of statements 1 to 36 to an individual in need.    -   40. The method of statement 39, wherein the disorder is        irritable bowel syndrome (IBS), constipation predominant IBS        (IBS-C), diarrhea predominant IBS (IBS-D) or diarrhea        predominant IBS (IBS-D).    -   41. The method of statement 39 or 40, wherein the pharmaceutical        formulation is administered subcutaneously.    -   42. The method of any one of statements 39 to 41, wherein the        pharmaceutical formulation is administered during acute pain        associated with IBS, IBS-C, IBS-D or IBS-M.    -   43. The method of statement 42, wherein the disorder is        diabetes, ischemia, reperfused tissue injury, dyslipidemia,        diabetic cardiomyopathy, myocardial infarction, acute coronary        syndrome, obesity, catabolic changes after surgery,        hyperglycemia, stroke, neurodegenerative disorders, memory and        learning disorders, islet cell transplant functional dyspepsia        and/or a disorder requiring regenerative therapy.    -   44. The method of any one of statements 39 to 43, wherein the        GLP-1 analogue is administered at a dose of from 50 μg to 150        μg.

REFERENCES

-   -   All patents, patent applications and publications referred to        throughout the application are incorporated herein by reference.

-   Hellström P M, et al., Clinical trial: the GLP-1 analogue ROSE-101    for management of acute pain in patients with irritable bowel    syndrome: a randomised, placebo-controlled, double-blind study.    Aliment Pharmacol Ther 29: 198-206, 2009

-   Michael Camilleri, et al., Effect of a glucagon-like peptide 1    analog, ROSE-010, on GI motor functions in female patients with    constipation-predominant irritable bowel syndrome. Am J Physiol    Gastrointest Liver Physiol 303: G120-G128, 2012

-   Mojsov, S., Int. J. Peptide Protein Research, 40:333-343 (1992)

-   Doyle, B. D. et al, Biophysical Signatures of Noncovalent Aggregates    Formed by a Glucagonlike Peptide-1 Analog: A Prototypical Example of    Biopharmaceutical Aggregation, Journal of Pharmaceutical Sciences,    vol. 94(12), p. 2749, 2005

-   U.S. Pat. No. 8,642,548 issued on Feb. 4, 2014, Peter Richardson et    al.,

-   U.S. Pat. No. 6,583,111

-   WO2007/028394

-   WO91/11457

-   Zölls, S et al., The AAPS Journal. Vol 15(4), p. 1200, 2013    (reference to MFI, page 78)

1. A liquid pharmaceutical formulation comprising a GLP-1 analoguehaving the amino acid sequence set forth in SEQ ID NO:3, wherein theliquid pharmaceutical formulation is at a pH of between 6.5±0.5 and7.5±0.5 and comprises a) histidine HCl at a concentration of about 10mM±10%, b) a sugar-based tonicity agent at a concentration of from 100mM to 450 mM c) polysorbate 20 at a concentration of from 0.005 to 0.05%and d) an amino acid excipient at a concentration of from 5 mM to 75 mM.2. The liquid pharmaceutical formulation of claim 1, wherein thesugar-based tonicity agent is mannitol at a concentration of from 150 mMto 350 mM.
 3. The liquid pharmaceutical formulation of claim 1, whereinthe sugar-based tonicity agent is sucrose at a concentration of from 145mM to 440 mM.
 4. The liquid pharmaceutical formulation of claim 1,wherein the amino acid excipient is methionine at a concentration offrom 5 mM to 25 mM.
 5. The liquid pharmaceutical formulation of claim 1,wherein the amino acid excipient is proline at a concentration of from25 mM to 75 mM.
 6. The liquid pharmaceutical formulation of claim 1,wherein the GLP-1 analogue is at a concentration of from 50 μg/ml to 2mg/ml.
 7. The liquid pharmaceutical formulation of claim 1, wherein theGLP-1 analogue is at a concentration of from 100 μg/ml to 1 mg/ml
 8. Theliquid pharmaceutical formulation of claim 1, wherein the GLP-1 analogueis at a concentration of from 100 μg/ml to 500 μg/ml.
 9. The liquidpharmaceutical formulation of claim 1, wherein the GLP-1 analogue is ata concentration of 300 μg/ml±20%.
 10. The liquid pharmaceuticalformulation of claim 1, wherein the surfactant is polysorbate
 20. 11.The liquid pharmaceutical formulation of claim 1, wherein the liquidpharmaceutical formulation is at pH 7.0±0.5.
 12. The liquidpharmaceutical formulation of claim 1, wherein the GLP-1 analoguecomprises the amino acid sequence set forth in SEQ ID NO:3 at aconcentration of from 50 μg/ml to 2 mg/ml, wherein the liquidpharmaceutical formulation comprises histidine-HCl at a concentration ofabout 10 mM, mannitol at a concentration of about 275 mM, methionine ata concentration of about 10 mM and polysorbate 20 at a concentration ofabout 0.02%.
 13. The liquid pharmaceutical formulation of claim 1,wherein the GLP-1 analogue comprises the amino acid sequence set forthin SEQ ID NO:3 at a concentration of from 50 μg/ml to 2 mg/ml, whereinthe liquid pharmaceutical formulation comprises histidine-HCl at aconcentration of about 10 mM, sucrose at a concentration of about 235mM, methionine at a concentration of about 10 mM and polysorbate 20 at aconcentration of about 0.02%.
 14. The liquid pharmaceutical formulationof claim 1, wherein the liquid pharmaceutical formulation is selectedfrom the group consisting of: a. a liquid pharmaceutical formulationcomprising a GLP-1 analogue having an amino acid sequence as set forthin SEQ ID NO:3, wherein the GLP-1 analogue is at a concentration of from100 μg/ml to 1 mg/ml and wherein the liquid pharmaceutical formulationcomprises histidine-HCl at a concentration of 10 mM, mannitol at aconcentration of 275 mM, methionine at a concentration of 10 mM andpolysorbate 20 at a concentration of 0.02%, and; b. a liquidpharmaceutical formulation comprising a GLP-1 analogue having an aminoacid sequence as set forth in SEQ ID NO:3, wherein the GLP-1 analogue isat a concentration of from 100 μg/ml to 1 mg/ml and wherein the liquidpharmaceutical formulation comprises histidine-HCl at a concentration of10 mM, sucrose at a concentration of 233 mM (8%), methionine at aconcentration of 10 mM and polysorbate 20 at a concentration of 0.02%.15. The liquid pharmaceutical formulation of claim 14, wherein theliquid pharmaceutical formulation is at pH 7.0±0.5.
 16. A single-dose ormultiple-dose container comprising the liquid pharmaceutical formulationof claim
 14. 17. A method of treating a disorder or condition in whichadministration of a GLP-1 or a GLP-1 analogue is indicated, the methodcomprising administering the liquid pharmaceutical formulation of claim14.
 18. The method of claim 17, wherein the disorder is irritable bowelsyndrome (IBS), constipation predominant IBS (IBS-C), diarrheapredominant IBS (IBS-D) or diarrhea predominant IBS (IBS-D).
 19. Themethod of claim 17, wherein the pharmaceutical formulation isadministered subcutaneously.
 20. The method of claim 18, wherein thepharmaceutical formulation is administered during acute pain associatedwith IBS, IBS-C, IBS-D or IBS-M.
 21. The method of claim 17, wherein thedisorder is diabetes, ischemia, reperfused tissue injury, dyslipidemia,diabetic cardiomyopathy, myocardial infarction, acute coronary syndrome,obesity, catabolic changes after surgery, hyperglycemia, stroke,neurodegenerative disorders, memory and learning disorders, islet celltransplant functional dyspepsia and/or a disorder requiring regenerativetherapy.
 22. The method of claim 17, wherein the GLP-1 analogue isadministered at a dose of from 50 μg to 150 μg.
 23. A method of treatinga disorder or condition in which administration of a GLP-1 or a GLP-1analogue is indicated, the method comprising administering the liquidpharmaceutical formulation of claim
 1. 24. The method of claim 23,wherein the disorder is irritable bowel syndrome (IBS), constipationpredominant IBS (IBS-C), diarrhea predominant IBS (IBS-D) or diarrheapredominant IBS (IBS-D).
 25. The method of claim 23, wherein thepharmaceutical formulation is administered subcutaneously.
 26. Themethod of claim 24, wherein the pharmaceutical formulation isadministered during acute pain associated with IBS, IBS-C, IBS-D orIBS-M.
 27. The method of claim 23, wherein the disorder is diabetes,ischemia, reperfused tissue injury, dyslipidemia, diabeticcardiomyopathy, myocardial infarction, acute coronary syndrome, obesity,catabolic changes after surgery, hyperglycemia, stroke,neurodegenerative disorders, memory and learning disorders, islet celltransplant functional dyspepsia and/or a disorder requiring regenerativetherapy.
 28. The method of claim 23, wherein the GLP-1 analogue isadministered at a dose of from 50 μg to 150 μg.
 29. A liquidpharmaceutical formulation comprising a GLP-1 analogue having an aminoacid sequence as set forth in SEQ ID NO:3, wherein the GLP-1 analogue isat a concentration of from 50 μg/ml to 2 mg/ml and wherein the liquidpharmaceutical formulation is at a pH of between of 6.5±0.5 and 7.5±0.5and comprises: a. histidine-HCl at a concentration of about 10 mM±10%,mannitol at a concentration of from 100 mM to 350 mM, and polysorbate 20at a concentration of from 0.01 to 0.03% (w/v); b. histidine-HCl at aconcentration of about 10 mM±10%, mannitol at a concentration of 165mM±10%, and polysorbate 20 at a concentration of from 0.01 to 0.03%(w/v); c. histidine-HCl at a concentration of 10 mM±10%, mannitol at aconcentration of 275 mM±10%, and polysorbate 20 at a concentration offrom 0.01 to 0.03% (w/v); d. histidine-HCl at a concentration of 10mM±10%, mannitol at a concentration of 275 mM±10%, polysorbate 20 at aconcentration of from 0.01 to 0.03% (w/v) and methionine at aconcentration of from 5 mM to 25 mM; e. histidine-HCl at a concentrationof 10 mM±10%, mannitol at a concentration of 165 mM±10%, polysorbate 20at a concentration of from 0.01 to 0.03% (w/v) and methionine at aconcentration of from 5 mM to 25 mM; f. histidine-HCl at a concentrationof 10 mM±10%, mannitol at a concentration of 275 mM±10%, polysorbate 20at a concentration of from 0.01 to 0.03% (w/v) and proline at aconcentration of from 25 mM to 75 mM; g. histidine-HCl at aconcentration of 10 mM±10%, mannitol at a concentration of 165 mM±10%,polysorbate 20 at a concentration of from 0.01 to 0.03% (w/v) andproline at a concentration of from 25 mM to 75 mM; h. histidine-HCl at aconcentration of 10 mM±10%, sucrose at a concentration of from 145 mM to450 mM, and polysorbate 20 at a concentration of from 0.01 to 0.03%(w/v); i. histidine-HCl at a concentration of about 10 mM±10%, sucroseat a concentration of about 235 mM±10%, and polysorbate 20 at aconcentration of from 0.01 to 0.03% (w/v); j. histidine-HCl at aconcentration of about 10 mM±10%, sucrose at a concentration of about290 mM±10%, and polysorbate 20 at a concentration of from 0.01 to 0.03%(w/v); k. histidine-HCl at a concentration of about 10 mM±10%, sucroseat a concentration of about 235 mM±10%, polysorbate 20 at aconcentration of from 0.01 to 0.03% (w/v) and methionine at aconcentration of from 5 mM to 25 mM; l. histidine-HCl at a concentrationof about 10 mM±10%, sucrose at a concentration of about 290 mM±10%,polysorbate 20 at a concentration of from 0.01 to 0.03% (w/v) andmethionine at a concentration of from 5 mM to 25 mM; m. histidine-HCl ata concentration of about 10 mM±10%, sucrose at a concentration of about235 mM±10%, polysorbate 20 at a concentration of from 0.01 to 0.03%(w/v) and proline at a concentration of from 25 mM to 75 mM, or; n.histidine-HCl at a concentration of about 10 mM±10%, sucrose at aconcentration of about 290 mM±10%, polysorbate 20 at a concentration offrom 0.01 to 0.03% (w/v) and proline at a concentration of from 25 mM to75 mM.
 30. A liquid pharmaceutical formulation comprising a GLP-1analogue having the amino acid sequence set forth in SEQ ID NO:3,wherein the liquid pharmaceutical formulation is at a pH of between6.5±0.5 and 7.5±0.5 and comprises histidine-HCl as buffering agent,sucrose or mannitol as tonicity agent, methionine or proline as anexcipient and a surfactant.